Dissasa et al.

BMC Veterinary Research (2022) 18:439

https://doi.org/10.1186/s12917-022-03508-w

RESEARCH Open Access

Isolation and identification of major

bacteria from three Ethiopian rift valley lakes
live and processed fish, and water samples:
implications in sanitary system of fish products
Guta Dissasa1*, Brook Lemma2 and Hassen Mamo3

Abstract
Bacterial pathogens are a great threat to fish production. Gram-negative bacteria are among the major bacterial fish
pathogens and are zoonotic with the potential to infect humans. This cross-sectional study was conducted to isolate
and identify major gram-negative bacteria from live and processed fish, and water samples from Lakes Hawassa, Lan-
ganoo and Ziway. A total of 674 different types of samples: 630 tissue samples (210 samples for each intestine, Kkid-
ney and liver collected from 210 live fish (Oreochromis niloticus, Cyprinus carpio and Clarias gariepinus), 20 processed
fish samples from lake Ziway fish processing center and 24 lake water samples were included in the study from each
lake. The mean values of pH, temperature, dissolved oxygen and nitrate in all water samples were within the normal
range at which most freshwater fish species become non-stressed. Of a total of 674 samples included in the study,
bacteria were isolated from 154(22.8%) samples with significant difference (P < 0.05) observed in some isolates with
respect to sample origin. Of these 154 isolates, 103(66.8%) isolates were gram-negative bacteria consisting of 15 spe-
cies based on morphology and a range of biochemical tests. From live fish samples, Escherichia coli was the dominant
species with 15 isolates followed by Edwardsiella tarda (12), Salmonella Paratyphi (10), Salmonella Typhi (9), Shigella
dysenteriae (7), Shigella flexneri (7), Klebsiella pneumonia (7), Enterobacter aerogenes (6), Enterobacter cloacae (5), Pseu-
domonas aeruginosa (5), Vibrio parahemolyticus (5), Aeromonas sobria (4), Citrobacter freundii (4), Citrobacter koseri (4)
and Plesiomonas shigelloides (3). The detection of the common fecal coliforms (E. coli, K. pneumoniae and E. aerogenes)
and Salmonella spp. in processed fish indicates the potential danger of passage of pathogenic bacteria and/or their
poisons to humans via infected and/or contaminated fish products. Human infection by pathogenic fish bacteria and
food poisoning is possible through contamination of fish product in fish production chain due to inadequate han-
dling, poor hygiene and contact with contaminated water. Therefore, producers, consumers and all other stakeholders
need to be cautious during handling, processing and consumption of fish harvested from the study lakes.
Keywords: Fish bacteria, Gram-negative bacteria, Physicochemical parameters, Rift-valley lakes

Background
Fish plays an important role in the human diet with
an ever-growing need globally. World fish production
has increased dramatically during the past 60 years, to
*Correspondence: [email protected] around 179 million tons in 2018 with a value of $401
1 billion and global fish consumption also increased from
Institute of Biotechnology, Addis Ababa University, Addis Ababa, Ethiopia
Full list of author information is available at the end of the article 9.0 kg per capita in 1961 to 20.5 kg in 2018 [1]. This is

© The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which
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Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 2 of 14

a tremendous change in the fishery industry. Ethiopia quality of the Ethiopian rift-valley lakes are scant. Hence,
depends on its inland lakes and rivers for fish produc- this study was aimed to assess the occurrence, distribu-
tion, and has over 200 edible fish species with annual fish tion, prevalence and identity of major gram-negative
production potential of about 94,500 thousand tons [2] enteric bacteria from live-caught and commercial pro-
although the potential remains much higher. cessed fish product of three common fish species—Nile
From a broader perspective, fish is among the top com- tilapia (Oreochromis niloticus), common carp (Cyprinus
ponents of aquatic biodiversity and is closely connected carpio) and catfish (Clarias gariepinus)—and water sam-
with several sectors of human socioeconomic life. How- ples from three Ethiopian rift-valley (inland) Lakes Lake
ever, fish health is a factor of aquatic ecosystem dynamics. Hawassa, Lake Langano and Lake Ziway in Ethiopia.
Pathogenic fish bacteria, among other variables, disturb
ecosystem dynamics and affect aquatic biodiversity and
Methods
fish wellbeing; erode the fishery industry and food secu-
The study area
rity. Moreover, fish consumption is associated with some
The study was on three Ethiopian rift-valley Lakes, Lake
serious bacterial infections mainly due to poor sanitary
Ziway, Lake Langano and Lake Hawassa, which are well-
facilities and practices around water systems, unhygienic
known for their common fish catches and tourist desti-
conditions in fishing and fish production chain. In par-
nations. These Lakes are some 163, 200 and 275 km to
ticular, gram-negative bacteria like Aeromonas spp., Fla-
the south of Addis Ababa respectively (Fig. 1). Morpho-
vobacterium spp., Pseudomonas spp., Edwardsiella spp.,
metric and other characteristics of the Lakes are shown
Vibrio spp., Acinetobacter spp. and Plesiomonas shigel-
in Supplementary Table 1 with information sources from
loides are a great threat to fish production [3, 4].
https://​latit​udelo​ngitu​de.​org/​et [11], World Lake Data-
Most of these bacterial fish pathogens are zoonotic
base [12], Wood and Talling 1988 [13].
with a potential to infect humans, and some of them are
serious [5]. Besides, fish pathogenic bacteria influence
fish population with substantial concerns for aquatic Sample size
biodiversity and fish industry, and food security. Stud- The sample size used to estimate the prevalence of bacte-
ies on fish microbial community provide a clue about rial infections in live fish samples was determined using
the hygienic status of the environment as water quality the formula n = Z 2 P(1 − P))/d 2 by Daniel and Cross
and fish diseases are closely linked [6]. Detection of fish (2013) [14], where ’n’ is the sample size, ’Z’ the confi-
pathogenic bacteria including novel ones, or change in dence interval (1.96), ’P’ the average prevalence estimate
the water microflora is an important indicator of envi- of 16.5% calculated from a review report in Ethiopia [15],
ronmental contamination. This is due to negative eco- and ’d’ the expected error (0.05).
logical changes in the natural fish habitat. Progressive
degradation of aquatic ecosystems is due to human activ- Study design and sampling strategy
ities associated with urbanization, industrialization, and The study design was cross-sectional and its period
agriculture resulting in the release of sewages of differ- was from February to August 2021. For sampling, the
ent nature and origin [7]. Additional factor that contrib- three Lakes were selected purposively that the Lakes are
utes to the emergence of novel fish pathogenic bacteria known for their common fish catches and from which
is unmanaged use of antibiotics and disinfectants [8]. fish are continuously harvested for subsistence and cash
Non-human factors like climate change and unpredict- [16]. Different sampling points of each Lake were used
able natural disasters seriously affect water bodies and all to measure physicochemical parameters, and collect live
life therein [9, 10]. fish and water samples.
The risk of contamination of water bodies, particularly Overall, 12 sampling sites, four from each Lake, were
inland closed systems like the Ethiopian rift-valley lakes, selected guided by the available information. The sam-
is particularly increasing due to clearly visible extensive pling points are considered major fishing grounds along
nearby development activities, urbanization and to cer- the shorelines. Accordingly, Amora Gedel (S1), Haile
tain extent frequent visits for various reasons including resort area (S2), Inlet of Tikurwuha River (S3) and Refer-
international ecotourism. Therefore, assessment of the ral Hospital area (S4) were sites of Lake Hawassa. Dole
physicochemical components of aquatic ecosystems and (S1), O’etu (S2), Wabishebelle (S3) and Yakona (S4) were
their bacterial community including inside resident fish sites of Lake Langano; and Abosa (S1), Bochessa (S2),
is essential to monitor fish health, product quality and Cafeteria (S3) and Wasiko (S4) were sites selected at Lake
related potential environmental and public health chal- Ziway. These sites were located at different directions of
lenges. Reports on bacterial pathogens in freshwater fish, the Lakes and were about 3-7 km apart. Thus, the sites
water physicochemical parameters and bacteriological were assumed to represent each lake.
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 3 of 14

Fig. 1 The study area

Water samples for nutrient and bacteriological analysis Fish samples

The on-spot-checkable water chemistry parameters The live fish samples were inspected for external abnor-
were tested in situ using routine methods [17]. Temper- malities, and were grouped into apparently healthy and
ature, electrical conductivity (EC) and dissolved oxygen clinically sick. The fish were euthanized by cervical dis-
(DO) were measured using temperature meter (YSI location in the fish’s normal water without using anes-
model 33 S-C-T meter, USA), conductivity meter (JEN- thesia. Trained individuals practiced the technique using
WAY, Multi-3410, UK) and multi parameter (Multi- appropriate equipment. Cervical dislocation is among the
3410, Germany) respectively. The pH was recorded methods recommended for fish sacrifice as it is relatively
using a digital pH meter (HI-99130, Italy). simple and effective for not big fish [21]. As our fish were
Surface water samples from each sampling point were small to medium in size, they were killed by inserting a
collected into sterile glass bottles (100 ml) as per stand- rod or thumb into the mouth, holding with the opposite
ard sampling procedure for lake or stream surface water hand and displacing it dorsally. Death was recognized by
chemistry for nutrient content testing [18] and bacte- cessation of movement, and confirmed by cessation of
riological analysis [19, 20]. The water samples were kept respiration (opercular movement) and cessation of heart-
dark, packed, and refrigerated. For live fish sampling, a beat (palpation); and finally by destruction of the brain.
random sampling technique was applied to represent The samples were separately packed in sterile plastic bags
the three target fish types (the tilapia, common carp and were shipped to Batu Fisheries and Other Aquatic
and catfish). In case of external disease occurrence, Life Research Center located at Batu town (formerly
diseased fish were purposively selected. The fish were Ziway), near Lake Ziway. The fish were dissected under
drawn from the peripheral, mid and central regions of aseptic conditions using a sterile dissecting scissor by fol-
the sampling points using fishing boats and gears of lowing established protocol [22, 23] and standard operat-
local fishers. ing procedures of bacteriology [19, 20].
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 4 of 14

Examination of the internal fish anatomy was made Biochemical characterization

by observing for any abnormalities including position, All the media used in the tests were from HIMEDIA,
size, color and other signs of damage. The digestive tract, India. The tests began with inoculating respective
gonads and visceral organs were removed by cutting the media with 24-h-old pure culture colonies. All incu-
esophagus and disconnecting them from the kidneys. bations were at 37 °C for 24 h and the expected color
Using sterile scalpel blade or forceps, tissue samples of changes confirmed test positivity. Briefly, indole pro-
the kidney, intestine and liver were aseptically transferred duction was tested by inoculating 10 ml of Dev trypto-
to sterile bottles (100 ml) with physiological saline, and phan broth, incubating and adding 2–3 drops of indole
were homogenized for bacteriological analysis [19]. reagent. Methyl red (MR) test was conducted by inocu-
Similarly, about 10 g muscle was cut from processed lating 10 ml of MR Voges-Proskauer (MR-VP) medium,
fish in Batu town fish processing center using a ster- incubating, and adding 2–3 drops of 0.05% MR. Voges-
ile knife, kept in sterile universal bottles (100 ml), and Proskauer (VP) test was done by inoculating 10 ml of
homogenized with 10 ml physiological saline solution. MR-VP medium, incubating, and adding 2–3 drops of 5%
From the homogenized samples of both live and pro- a-nephtol followed by 40% of KOH and shaking and leav-
cessed fish, 1 ml aliquot was drawn and further homog- ing it open for an hour.
enized in a clean, dry sterile beaker containing 9 ml of For catalase test, a small amount of bacterial colony was
distilled water to have 1:10 dilution [20]. transferred to clean glass slide using a sterile loop and a
Finally, all the three sample types (water, live fish and drop of hydrogen peroxide was added, and the formation
processed fish) were packaged and transported on ice to of bubbles was checked for. For citrate utilization test,
Health Biotechnology Laboratory of Institute of Biotech- Simmons citrate agar slant was inoculated and incubated.
nology, Addis Ababa University, and were stored at 4 °C Hydrogen sulfide or triple sugar iron (TSI) test was done
for nutrient and bacteriological analysis. by inoculating TSI by first stabbing through the center of
the medium to the bottom of the tube and then streaking
Water nutrient content analysis the surface of the agar slant, and incubating. Similarly,
The nitrate content of the transported water samples urea production was tested using Christensen’s Urea
was determined using the sodium salicylate method and Agar slant. Sugar fermentation test was conducted using
phosphate by ascorbic acid method using UV–visible sugar broth medium prepared by mixing 1 g peptone,
spectrophotometer (Lambda, CE1021, and Australia). 0.3 g meat extract, 0.5 g table salt, 0.5 g sugar and 0.008 g
phenol indictor in 100 ml distilled water. Three tubes
Bacteria culture and colony morphology having three different sugars (glucose, sucrose, lactose) in
All bacteriological experiments were performed follow- the broth medium were inoculated, and incubated.
ing Society of American Bacteriologists Manual of micro-
biological methods [19], Bergey’s manual of determinative Data analysis
bacteriology [20] and the respective media manufactur- Bacterial infection status of the different sample types
er’s instructions. It was under complete aseptic condi- was determined and the proportion of infected samples/
tions. All incubations were for 24 h at 37 °C. isolates was compared between various categories using
First, sample swabs were spread or streaked across the Chi-squared test. Bacterial species diversity in differ-
nutrient agar medium (Oxoid, England) and incubated ent sample types was compared using one-way analysis
under aerobic condition. Then, specific colonies were of variance (ANOVA). Statistical analysis was performed
picked-up and inoculated on selective and differential using IBM SPSS software version 20 (IBM, Chicago,
media Xylose Lysine Deoxycholate (XLD) agar (HIME- USA) and p < 0.05 was considered statistically significant.
DIA, India), and incubated further. Suspected bacterial
colonies were picked-up, inoculated into tryptone soy Results
broth (HIMEDIA, India), and incubated. Colony mor- Sample type and total number
phology like form, elevation, margin, surface and pig- The sampled live-caught fish were 210. Of these, 42(20%)
mentation were examined. Colony color was determined had evident external health problems and the rest were
by visual inspection of bacterial cell suspensions using apparently healthy both externally and after dissection.
fresh culture. The colonies were screened microscopi- In terms of species, 105 were tilapia (O. niloticus), 60
cally using simple and differential (gram) staining. The common carp (C. carpio) and 45 catfish (C. sgariepinus).
morphologically presumptively identified isolates were Overall, 90, 50 and 70 of the fish were from Lake Ziway,
stored at -20 °C in 50% glycerol (Fine Chemical, Ethiopia) Langanoo and Hawassa respectively. The diseased fish
using 1.8 ml cryovials (IMEC, China) for biochemical were 23 tilapia (8 from Lake Hawassa, 2 from Langanoo
identification. and 13 from Ziway) plus 19 carps (12 from Lake Ziway
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 5 of 14

and 7 from Hawassa). Intestine, kidney and liver tissue While all of the above species have been isolated from
samples were taken from each fish making the number live fish samples, E. aerogenes, E. cloacae, P. shigelloides
of samples of live fish origin 630. The size of processed and S. typhi were not detected in any of the water sam-
fish samples was 20. The total number of water samples ples. Only E. coli, K. pneumoniae and E. aerogenes (the
was 36. That is, duplicate samples from each 4 sampling common fecal coliforms) and Salmonella spp. were
points of each lake making 24 samples for bacteriologi- detected in processed fish samples. E. coli, K. pneumo-
cal analysis, and additional 12 samples from each point of nia and S. Paratyphi were detected in all the three sam-
each lake for water chemistry. Hence, the overall sample ple types. Bacteria isolated from water samples, but not
number undergone bacteriological analysis was 674. from processed fish, were A. sobria, C. freundii, C. koseri,
E. tarda, P. aeruginosa, S. dysenteriae, S. flexneri and V.
Physicochemical parameters parahemolyticus. Conversely, E. aerogenes and S. typhi
The mean pH values of the three Lakes were nearly simi- were identified from processed fish but not from water
lar ranging 7.91–8.68. The temperature of Lake Hawassa samples (Table 1). The distribution of individual bacte-
was higher (25.8 °C) than that of the other two that had rial species was not statistically significant with respect
the same measure. Lake Ziway had the lowest OD (3.93 to sample type.
± 0.18 mg/ml) and Hawassa the highest (6.27 ± 0.90 mg/
ml). The highest EC (µS/cm) was that of Lake Hawassa Bacteria distribution by live fish tissue
(1673.5 ± 0.48) followed by Langanoo (1447 ± 5.70) The highest number of isolates (37(17.6%)) were recov-
and the least Ziway (289.5 ± 14). It is notable that the ered from the intestine followed by the liver (26(12.4%))
EC of Lake Ziway was fivefold below that of Hawassa. and kidney 19(9.1%) with statistically significant differ-
All records including for nitrate were highest in Lake ence (p < 0.0001). The most frequently isolated bacte-
Hawassa and least in Ziway except for pH. The highest rium from live fish was E. coli with 5 isolates (2.4%) in the
mean phosphate level was for Lake Langano followed by intestine and 5(3.6%) in the liver (Table 2). E. aerogenes
Hawassa and Ziway with all the three lakes having higher was the most frequent isolate from the kidney (2.1%). The
values than the standard limit. However, all the measured least frequently isolated bacterium from live fish was P.
physicochemical parameters of the three Lakes (Supple- shigelloides (1.4%). There was no significant difference
mentary Table 2) were at optimum level or at least within
the tolerance range of most freshwater fish species.
Table 1 The number and proportion of samples positive for
Bacterial isolates Gram-negative bacteria species among water (N = 24) and live
From the 674 samples, 154(22.8%) were positive for fish (N = 210) samples collected from Lakes Hawassa, Langanoo
bacteria (51 g-positive and 103 g-negative isolates). The and Ziway, and processed fish samples (N = 210) from Batu town
gram-negative bacteria were further tested and identi- Bacteria spp Sample type (n = 154)
fied into 15 species. All isolates were short rods and non- Water, n(%) Live fish, n(%) Processed Total
spore formers (Supplementary Fig. 1). Colonies of the fish, n(%)
isolates had different form, elevation, margin, surface,
A. sobria 1(5.9) 3(2.4) 0(0.0) 4(2.6)
colour and optical characteristics (Supplementary Fig. 2).
C. freundii 1(5.9) 3(2.4) 0(0.0) 4(2.6)
The prevalence of gram-negative bacteria among
C. koseri 1(5.9) 3(2.4) 0(0.0) 4(2.6)
water, processed fish and live-caught fish samples was
E. tarda 2(11.8) 10(8.1) 0(0.0) 12(7.8)
50% (12/24), 45% (9/20) and 39% (82/210) respectively.
The difference was statistically significant (p < 0.0001). E. aerogenes 0(0.0) 4(3.2) 1(7.7) 5(3.2)
Escherichia coli was the dominant species with 15 iso- E. cloacae 0(0.0) 5(4.0) 0(0.0) 5(3.2)
lates (12 from live-caught fish, 2 processed fish, 1 water E. coli 1(5.9) 12(9.7) 2(15.4) 15(9.7)
sample) followed by 12 Edwardsiella tarda (10 live fish, K. pneumoniae 1(5.9) 4(3.2) 2(15.4) 7(4.5)
0 processed fish, 2 water sample)), 10 Salmonella Para- P. shigelloides 0(0.0) 3(2.4) 0(0.0) 3(1.9)
typhi (7 live fish, 2 processed fish, 1 water sample) and P. aeruginosa 1(5.9) 4(3.2) 0(0.0) 5(3.2)
9 Salmonella typhi (7 live fish, 2 processed fish, 0 water S. paratyphi 1(5.9) 7(5.6) 2(15.4) 10(6.5)
sample). The other gram-negative bacteria were Shigella S. typhi 0(0.0) 7(5.6) 2(15.4) 9(5.8)
dysenteriae, Shigella flexneri, Klebsiella pneumonia, S. dysenteriae 1(5.9) 6(4.8) 0(0.0) 7(4.5)
Enterobacter cloacae, Pseudomonas aeruginosa, Vibrio S. flexneri 1(5.9) 6(4.8) 0(0.0) 7(4.5)
parahemolyticus, Aeromonas sobria, Citrobacter freun- V. parahemo- 1(5.9) 4(3.2) 0(0.0) 5(3.2)
lyticus
dii, Citrobacter koseri, Enterobacter aerogenes and Plesio-
Total 12(50) 82(39.1) 9(45) < 0.0001
monas shigelloides.
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 6 of 14

Table 2 Distribution of bacteria in the kidney, liver and intestine of live fish caught from Lakes Hawassa, Langanoo and Ziway (N =
210)
Isolated bacteria Intestine, n(%) Kidney, n(%) Liver, n(%) Total, n(%) P-value

A. sobria 1(0.5) - 2(1.4) 3(1.4) 0.064

C. freundii 2(0.9) - 1(0.5) 3(1.4) 0.244
C. koseri 2(0.9) 1(0.5) - 3(1.4) 0.064
E. tarda 4(1.9) 2(0.9) 4(1.9) 10(4.8) 0.332
E. aerogenes 1(0.5) 3(2.1) 1(0.5) 5(2.4) 0.394
E. cloacae+ 2(0.9) 2(0.9) 1(0.5) 5(2.4) 0.630
E. coli* 5(2.4) 2(0.9) 5(3.6) 12(5.7) 0.104
K. pneumoniae 1(0.5) 2(0.9) 1(0.5) 4(1.9) 0.579
P. shigelloides 1(0.5) 2(0.9) - 3(1.4) 0.244
P. aeruginosa 2(0.9) 1(0.5) 1(0.5) 4(1.9) 0.702
S. paraTyphi 3(1.4) 2(0.9) 2(0.9) 7(3.3) 0.770
S. Typhi 4(1.9) 1(0.5) 2(0.9) 7(3.3) 0.406
S. dysenteriae 3(1.4) - 3(1.4) 6(2.9) 0.262
S. flexneri 3(1.4) - 3(1.4) 6(2.9) 0.166
V. parahemolyticus 3(1.4) 1(0.5) - 4(1.9) 0.048*
Total 37(17.6) 19(9.1) 26(12.4) 82(39.1) < 0.0001*
As Aeromonas sobria, Cf Citrobacter freundii, Ck Citrobacter koseri, Ct Edwardsiella tarda, Ea Enterobacter aerogenes, Ec+ Enterobacter cloacae, Ec*i Escherichia coli, Kp
Klebsiella pneumonia, Ps Plesiomonas shigelloides, Pa Pseudomonas aeruginosa, Sp Salmonella Paratyphi, St Salmonella typhi, Sd Shigella dysenteriae, Sf Shigella flexneri,
Vp Vibrio parahemolyticus
*
means significant at 5% level

in the distribution of individual bacterial species in the Bacteria distribution among clinically sick fish
three fish tissues except for V. parahemolyticus whose Out of 42 clinically diseased tilapia and catfish samples
prevalence in the intestine was significantly higher (p = from the three lakes, 36(85.7%) were positive for bacterial
0.048). infection. These were 19 tilapias (11 from Lake Hawassa
and 8 from Lake Ziway), and 17 carps (8 from Lake
Bacteria distribution by fish species Hawassa and 9 from lake Ziway). No bacteria were iso-
Except A. sobria, all other isolates were recovered from lated from diseased fish caught from Lake Langano (Sup-
tilapia (O. niloticus) live fish catches. Similarly, all other plementary Table 3). E. tarda was isolated from 5 tilapias
bacterial species were isolated from catfish (C. gariepi- and 2 carps in Lake Hawassa with overall prevalence of
nus) with the exception of P. shigelloides. However, C. 19.4% which was significantly higher than the occurrence
freundii, C, koseri and V. parahemolyticus were not iso- of any other species among sick fish from both Lakes (p
lated from carps. Significant variation (p < 0.0001) was = 0.046).
recorded regarding the prevalence of bacterial infection
in the three fish species (Table 3). Bacteria distribution in live fish by sampling site
Majority of Lake Hawassa isolates were from S1 (22.9%)
Bacteria distribution in live fish from the three lakes and S4 (18.6%). In Lake Langano, most isolates were from
The highest number of isolates were from Lake Hawassa S2 (10%). Similarly, most of the isolates were from sam-
(34(48.6%)) followed by Ziway (35(43.8%)) and Langanoo pling points S3 (18.8%) and S1 (13.8%) of Lake Ziway. The
the least with only 13(21.7%). The prevalence of bacterial distribution of A. sobria, C. freundii, C. koseri, E. cloa-
infection in the three Lakes was statistically significant (p cae, E. coli, P. aeruginosa and V. parahemolyticus varied
< 0.0001). All the fifteen bacterial species (100%) isolated significantly across live fish sampling sites of each Lake
in this study were in Lake Hawassa fish samples. Simi- (Table 5).
larly, all of the species were also isolated from Lake Ziway
fish samples except C. freundii. On the other hand, only Bacteria from water samples
9(60.0%) of the 15 species were detected from Lake Lan- The prevalence of bacteria in water samples from Lake
gano fish samples (Table 4). The data shows the highest Hawassa was 50%, Ziway 33.3% and Langanoo 16.7%
bacterial species composition in Lake Hawassa and the with significant difference (p = 0.046). A. sobria, C.
least in Langanoo. koseri, S. paratyphi, S. flexneri and V. parahemolyticus
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 7 of 14

Table 3 Number (percentage) of bacterial species isolated from three fish species caught from Lakes Hawassa, Langanoo and Ziway
Isolated bacteria Carps (Cc) N = 60 Tilapia (On) N = 105 Catfish (Cg) N = 45 Total n = 210 P-value

A. sobria 2(3.3) - 1(2.2) 3(1.4) 0.244

C. freundii - 2(1.9) 1(2.2) 3(1.4) 0.064
C. koseri - 1(1.4) 2(4.4) 3(1.4) 0.244
E. tarda 3(5) 4(3.8) 3(6.7) 10(4.8) 0.770
E. aerogenes 1(1.7) 1(1.4) 3(6.7) 5(2.4) 0.216
E. cloacae+ 1(1.7) 1(1.4) 3(6.7) 5(2.4) 0.079
E. coli* 3(5) 6(5.7) 3(6.7) 12(5.7) 0.084
K. pneumoniae 1(1.7) 2(1.9) 1(2.2) 4(1.9) 0.579
P. shigelloides 2(3.3) 1(1.4) - 3(1.4) 0.244
P. aeruginosa 1(1.7) 2(1.9) 1(2.2) 4(1.9) 0.702
S. paratyphi 2(3.3) 3(2.9) 2(4.4) 7(3.3) 0.422
S. typhi 2(3.3) 3(2.9) 2(4.4) 7(3.3) 0.579
S. dysenteriae 2(3.3) 3(2.9) 1(2.2) 6(2.9) 0.422
S. flexneri 3(5) 1(1.4) 2(4.4) 6(2.9) 0.125
V. parahemolyticus - 2(1.9) 2(4.4) 4(1.9) 0.125
Total 23(38.3) 32(30.5) 27(60.0) 82(39.1) < 0.0001**
**
On O. niloticus, Cg C. gariepinus, Cc C. carpio means significant at 5% level, As Aeromonas sobria, Cf Citrobacter freundii, Ck Citrobacter koseri, Ct Edwardsiella tarda, Ea
Enterobacter aerogenes, Ec+ Enterobacter cloacae, Ec* Escherichia coli, Kp Klebsiella pneumonia, Ps Plesiomonas shigelloides, Pa Pseudomonas aeruginosa, Sp Salmonella
ParaTyphi, St Salmonella Typhi, Sd Shigella dysenteriae, Sf Shigella flexneri, Vp Vibrio parahemolyticus

Table 4 Number (percentage) of bacteria isolated from live-caught fish samples from Lakes Hawassa, Langanoo and Ziway
Isolated bacteria Lake Hawassa N = 70 Lake Langano N = 60 Lake Ziway N = 80 Total N = 210 P-value

A. sobria 2(2.9) - 1(1.3) 3(1.4) 0.064

C. freundii 2(2.9) 1(1.7) - 3(1.4) 0.187
C. koseri 2(2.9) - 1(1.3) 3(1.4) 0.187
E. tarda 5(7.1) 2(3.3) 3(3.8) 10(4.8) 0.296
E. aerogenes 1(1.4) 1(1.7) 3(3.8) 5(2.4) 0.394
E. cloacae+ 2(2.9) - 3(3.8) 5(2.4) 0.098
E. coli* 5(7.1) 2(3.3) 5(6.3) 12(5.7) 0.146
K. pneumoniae 2(2.9) - 2(2.5) 4(1.9) 0.125
P. shigelloides 1(1.4) 1(1.7) 2(2.5) 4(1.9) 0.579
P. aeruginosa 1(1.4) - 2(2.5) 3(1.4) 0.302
S. paratyphi 3(4.3) 1(1.7) 3(3.8) 7(3.3) 0.296
S. typhi 2(2.9) 3(5) 2(2.5) 7(3.3) 0.729
S. dysenteriae 2(2.9) 1(1.7) 3(3.8) 6(2.9) 0.579
S. flexneri 3(4.3) 1(1.7) 2(2.5) 6(2.9) 0.536
V. parahemolyticus 1(1.4) - 3(3.8) 4(1.9) 0.072
Total 34(48.6) 13(21.7) 35(43.8) 82(39.1) < 0.0001**
As Aeromonas sobria, Cf Citrobacter freundii, Ck Citrobacter koseri, Ct Edwardsiella tarda, Ea Enterobacter aerogenes, Ec+ Enterobacter cloacae, Ec* Escherichia coli, Kp
Klebsiella pneumonia, Ps Plesiomonas shigelloides, Pa Pseudomonas aeruginosa, Sp Salmonella ParaTyphi, St Salmonella Typhi, Sd Shigella dysenteriae, Sf Shigella
flexneri, Vp Vibrio parahemolyticus
**
means significant at 5% level

were detected in Lake Hawassa. The only bacteria were exclusive to it (Supplementary Table 4). Not only
detected in Lake Langano and exclusive to it were C. in terms of prevalence but bacterial species diversity as
freundii and K. pneumonia. Similarly, from 4 species well, Lake Hawassa was the most diverse and Langa-
in Lake Ziway, 3 (E. coli, P. aeruginosa, S. dysenteriae) noo the least.
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 8 of 14

Table 5 Number (percentage) of bacteria isolated from fish sampled from four different sites (S1-S4) of Lakes Hawassa, Langanoo and
Ziway
Isolated bacteria Lake Hawassa n = 70 Lake Langano n = 60 Lake Ziway n = 80 Total n = 210 P-value
S1 S2 S3 S4 S1 S2 S3 S4 S1 S2 S3 S4

A. sobria 1(1.4) - - 1(1.4) - - - - - - 1(1.3) - 3(1.4) 0.001*

C. freundii 2(2.9) - - - - - 1(1.7) - - - - - 3(1.4) 0.001*
C. koseri 2(2.9) - - - - - - - 1(1.3) - - - 3(1.4) 0.001*
E. tarda 3(4.3) - - 2(2.9) - 2(3.3) - - - 2(2.5) 1(1.3) 10(4.8) 0.096
E. aerogenes - - - 1(1.4) - - 1(1.6) - 1(1.3) 1(1.3) - 1(1.3) 5(2.4) 0.206
E. cloacae 1(1.4) - 1(1.4) - - - - - 1(1.3) 1(1.3) 1(1.2) - 5(2.4) 0.003*
E. coli 2(2.9) 1(1.4) - 2(2.9) 1(1.7) - - 1(1.7) 2(2.5) - 1(1.3) 2(2.5) 12(5.7) 0.016*
K. pneumoniae - 1(1.4) - 1(1.4) - - - - 1(1.3) - 1(1.3) 4(1.9) 0.153
P. shigelloides - - - 1(1.4) - 1(1.7) - - 2(2.5) - - 3(1.4) 0.287
P. aeroginosa 1(1.4) - - - - - - - - - 2(2.5) - 4(1.9) 0.003*
S. paratyphi 2(2.9) 1(1.4) - - - - 1(1.7) - 2(2.5) - 1(1.3) - 7(3.3) 0.364
S. typhi - 1(1.4) - 1(1.4) 1(1.7) 1(1.7) 1(1.7) 1(1.3) 1(1.3) - - 7(3.3) 0.068
S. dysenteriae 1(1.4) - - 1(1.4) - 1(1.7) - - 1(1.3) - 2(2.5) - 6(2.9) 0.413
S. flexneri - 1(1.4) - 2(2.9) - 1(1.7) - - - - 2(2.5) - 6(2.9) 0.158
V. parahemo - - - 1(1.4) - - - - - - 2(2.5) 1(1.3) 4(1.9) 0.007*
Total 15(21.4) 5(7.1) 1(1.4) 13(18.6) 2(3.3) 6(10) 3(5) 2(3.3) 12(15) 3(3.8) 15(18.8) 5(6.3) 82(39.1)
S1 sample site 1, S2 sample site 2, S3 sample site 3, S4 sample site 4, n number of isolates
*
statistically significant;—not isolated, V. parahem, V. parahemolyticus

Discussion The mean DO values of 6.27 mg/l (Lake Hawassa),

The mean values of all the measured physicochemical 5.10 mg/l (Lake Langano) and 3.93 mg/l (Lake Ziway)
parameters in the three lakes were within the standard indicate relatively better aeration at lakes Hawassa and
limit for fish health [24–29]. In light of this, the lakes Langanoo than Ziway at least during the study period and
could be considered fitting for fish survival. Water qual- sampling time. This might be attributable to variations in
ity factors such as DO, temperature, ammonia, phos- temperature and flow rate in the Lakes during the sam-
phate, pH, alkalinity, hardness and clarity affect fish pling season as lower temperature and good flow rate are
health in multiple ways. Each water quality factor inter- associated with higher ODs [24, 30]. Most DO in ponds
acts with and influences other parameters, sometimes is produced during photosynthesis by aquatic plants
in complex ways and what may be fatally toxic in one and algae. For this reason, DO increases during daylight
situation can be harmless in another. Nevertheless, we hours, declines during the night, and is lowest just before
did not test many other parameters, which are impor- daybreak. DO concentrations below 5 mg/l may be harm-
tant water quality factors for logistics reasons. The ful to fish and piping (gulping air at the surface) may be
effect of sampling site, season and hour on the param- observed when DO falls below 2 mg/l. Low levels of DO
eters is usually considerable. For instance, DO values are most frequently associated with hot, cloudy weather,
significantly vary when the sampling site is surface algae die-offs, or heavy thunderstorms [30]. Overall, poor
water and along shorelines compared to deep water, or water quality is a key factor for low fish yields. In a pond
at the center. Our sampling was on surface water. study, whereas increase in temperature and OD was cor-
The oxygen requirements of fish also depend on a related with tilapia growth rate, increase in conductivity
number of other factors including temperature, pH, and pH showed the opposite [31]. Different fish species
and ­CO2 level of the water, as aforementioned, and the have different requirements for water DO concentration
metabolic rate of the fish. Therefore, changes in these [27], and water temperature may influence fish feeding,
physicochemical parameters in the aquatic environ- growth and overall behavior.
ment are primary causes of fish stress [27] although the Similarly, the data showed insignificant pollution by
health impact of such stress may depend not only on nitrogenous wastes at the sampling time although ammo-
the severity of the stress, but on its duration and the nia is a pollutant frequently found in aquatic ecosystems.
fish’s overall physiological status. In fish, ammonia can cause physical damage, alter its
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 9 of 14

behavior such as lower swimming activity and feeding A study on marine fish found significantly higher poten-
behavior, and oxidative stress response, and even cause tial pathogenic bacteria in kidneys than in liver samples
death. Exposure to ammonia also increases fish physi- and a variation was found between the fish species [39].
ological stress and recent evidence suggested that once The authors reported that significant differences were
exposed, fish suffer from reduced antioxidant defenses observed between fish species, organs and sites, indicat-
and thus increased oxidative tissue damage even if water ing the importance of the environmental conditions on
quality was improved [32]. the fish microbiome. Although we did not investigate
The only exception was phosphate level in Lake Lan- bacteria in different portions of the fish gut, it appears
gano which was relatively high (2.34–4.48 mg/l). The that microenvironment dynamics along the gut of vari-
highest concentration of phosphate (4.48 mg/l) recorded ous fish species may influence the composition and abun-
for Lake Langano O’etu site (S2) was higher than the dance of the bacterial flora [40].
standard limit although the mean value was normal. The In African catfish, higher bacterial load was recovered
mean value of phosphate for Langanoo was substan- from the intestine than other organs like skin and gills
tially higher than that of Hawassa, and that of Ziway was [41]. There is evidence that dense microbial popula-
much lower. In the early 1990s, it was reported that Lakes tions occur within the intestinal contents, with numbers
Ziway and Hawassa (then Hawassa) were phosphorus- of bacteria much higher than those in the surrounding
limited, whereas Langanoo has surplus phosphorus [33]. water, like the current finding, indicating that the intes-
It appeared that Lake Langano remained phosphate sur- tines provide favorable ecological niches for these organ-
plus. In the 1960s, however, increases in phosphorus in isms [42]. However, the method of intestinal bacteria
the lower Lakes raised considerable public concern [34]. sampling varied in different surveys. Some used anal
The augment in phosphorus could be due to increased swabs, others only the intestinal contents, while still in
surface runoff from phosphate containing fertilizers others intestinal tract and contents were homogenized
and certain industrial wastes. Phosphate is an essential and used for culturing [43]. Austin and Al-Zahrani (1988)
plant nutrient. However, high phosphate level can lead [44] distinguished between the flora of the gut contents
to eutrophication boosting plant/algae growth. These and that intimately associated with the wall of the gastro-
plants/algae eventually decay and cause DO depletion intestinal tract, and noted that scanning electron micros-
in the water threatening certain species of fish favoring copy showed only sparse microbial colonization of the
phosphate-pollution-tolerant organisms. wall. Although the genera present in the gut generally
Although it appeared that the physicochemical param- seem to be those from the environment or diet which can
eters of the Lakes were normal, the proportion (20%) of survive and multiply in the intestinal tract, there is evi-
fish with visible health problems is not a good sign. Alter- dence for a distinct intestinal microflora in some species
natively, it may be plausibly argued that encountering [43]. This same author reviewed the progressive decline
relatively lesser number of clinically diseased fish with in the numbers of aerobic heterotrophic bacteria along
visible pathological changes might be attributed to the the digestive tract. Anaerobes were detected only in the
good water quality of the Lakes during the study period upper intestine and in the intestinal contents.
notwithstanding the seasonal fluctuations in water qual- However, other investigators [45] found that numbers
ity [35, 36]. At least during the study season, majority of bacteria in freshwater salmonids increased between
of the fish caught were less stressed. The effect of stress the stomach and the posterior portion of the intestine.
on freshwater fish may be a factor of the severity of the The authors suggested that these numbers must repre-
stress, its duration and the overall physiological state of sent active multiplication in the tract, as they could not
the fish [37]. be accounted for by ingestion. The numbers detected
The proportion of bacterial isolates from the intestine in this survey were probably an artificially low estimate
of live-caught fish (17.6%) was significantly higher than since the methods used did not allow for the isolation
that of the liver (12.4%) or kidney (9.1%). But, there were and growth of strict anaerobes, species sensitive to oxy-
slight variations with respect to the individual bacteria gen, nutritionally fastidious species, or those requir-
species. For instance, the number of isolates from live- ing low growth temperatures (< 20 °C). In addition, the
caught fish intestine was equal to that of the liver con- counts obtained were based on the total tissue weight
cerning E. coli which was the most prevalent bacterium in each sample, while the bacteria actually populate
isolated. The second most prevalent in this study, E. only the epithelial surface of the tract, and the rest of
tarda, was similarly isolated from the intestine and liver the tissue sterile. There was no significant difference
in equal proportions. E. tarda is a cause of rare but fatal in the bacterial flora of fish of different species, sex,
food-/waterborne infection in man [38]. Contrarily, E. breeding status, weight, or geographical source. But,
aerogenes was most frequently isolated from the kidney. microenvironment characteristics at various locations
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 10 of 14

through the gastrointestinal tract of fish influence the Moreover, the fact that most of E. coli isolated during
composition and abundance of gut bacteria [46] and this study was from fish intestine reflecst warm-blooded
bacterial counts significantly differed between species, animal pollution level of the water. E. coli can have a
sources and feeding habits of examined fishes [47]. long-term survival and can multiply depending on fish
Fish internal organs such as the spleen, liver, and kid- and water temperatures [64–67]. It is known that fish
neys are expected to be sterile [48]. Nonetheless, evi- possess distinct intestinal microbiota, but nutritional
dence accumulates for bacteria from internal organs status or feeding habits, trophic level, species (attribute
of apparently healthy fish [49] in agreement with the to complexity of the fish digestive system) and the envi-
current study. Possible fish immune compromise may ronmental conditions (salinity of the habitats and the
explain such observations. Immune defect could hap- bacterial load in the water) are the most influential fac-
pen due to stress. Fish stress could be associated with tors which change the intestinal microbiota composition
poor water quality, temperature changes, nutritional and abundance [57]. This may explain the observed dif-
deficiencies, overcrowding, trauma, parasitism, pri- ferences in the distribution of bacteria from fish samples
mary viral infections [50–52], among others. For in the three Lakes. While bacteria in water can influence
instance, fish immunity was found significantly affected the microbial flora associated with fish, the reverse is also
by lower temperatures [53] making them susceptible to worth consideration.
obligate or facultative pathogenic bacteria such as A. Shigella spp. and Salmonella spp. are pathogenic bac-
hydrophila. teria found in animal, human or environmental reservoir.
The bacterial flora of the gut of two marine fish has Although contamination of fish products with these bac-
been investigated in an attempt to clarify the relation- teria is commonly from the environment, their incidence
ship between these bacteria and the bacterial flora of in ready-to-eat fish product due to unhygienic handling
their diets, and to determine the effect of the degree of cannot be ruled out [68]. Citrobacter, Enterobacter and
specialization of the digestive tracts on their floras [54]. Klebsiella are indigenous to general environment and
Bacterial flora of fish with relatively undeveloped diges- frequently present in fish but most of these bacteria are
tive tracts reflected that of the fishes’ food, whereas fish considered non-pathogenic environmental strains. The
with more specialized tracts have a distinctive gut micro- bacterial species isolated from processed fish (E. coli, K.
flora. In the red sea bream, the composition of the bacte- pneumonia and S. paraTyphi) were also recovered from
rial floras of the stomach and intestine changed with time the water samples.
after feeding. Half an hour after ingestion, most of the The variation in the number of isolates and bacterial
bacteria isolated from the stomachs resembled those of species between sampling sites of the study Lakes might
the fish meat diet, but after 6 h, vibrios resistant to bile be attributed to the relative distance and degree of expo-
and low pH predominated. Further work comparing rep- sure to the nearby point source pollution around the
resentative strains of these indigenous vibrios from the study area. Disruption of the environmental microbiome
red sea bream with other isolates from the stomach and after an earthquake followed by seasonal variation in the
intestine showed that the vibrios were able to survive in water quality was noted although restoration of these
the presence of gastric juice at pH 4, and were able to microbial communities as a function of time and sanita-
grow, although at a reduced rate, at pH 5, while most of tion practices occurred in Nepal [69].
the other isolates were inhibited by these conditions [55]. All positive live fish samples were positive for at least
In this study, the water samples were positive for A. one isolate of all the 15 bacteria species recovered. This
sobria, Citrobacter spp., E. tarda, E. coli, K. pneumo- shows the higher bacteria species diversity in fish com-
niae, P. aeruginosa, S. typhi, S. dysenteriae and S. flexneri. pared to the aquatic environment wherein only 4 species,
Detection of these and other related bacteria both in E. aerogenes, E. cloacae, P. shigelloides and S. typhi, were
marine and freshwater habitats and fish has been widely characterized although the water bacteria prevalence was
recorded [56–59]. Aeromonas is associated with a range higher. The finding suggests that the bacteria detected
of human opportunistic infections including enteritis in fish internal tissue might constitute the natural fish
and septicemia [60–62] and is one of the most common microbiota and/or the fish bacteria source might be their
pathogens in tropical fish [63]. E. coli was the most fre- diet. Alternatively, the fish might have been exposed to
quently detected bacterium in processed and live-caught seasonal or occasional biological pollutants which have
fish samples in this study. Total and fecal coliforms such been diluted or neutralized from the environment. Such
as E. coli are indicators of fecal contamination of aquatic dynamics in the aquatic ecosystem may explain par-
environments and food. Detection of E. coli in processed ticularly the absence of human urinary and respiratory
fish samples could be due to unhygienic handling during tracts pathogens E. aerogenes and E. cloacae [70] in water
processing. samples and their detection in fish. Another surprising
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 11 of 14

finding is the absence of P. shigelloides in the water sam- were isolated from the water samples, were also recov-
ples. The common environmental reservoirs for this ered from fish in the Lake. Moreover, an outbreak of A.
organism include freshwater ecosystems and estuaries hydrophila associated with a certain parasite in pond of
and inhabitants of these aquatic environs, and a series of African catfish fingerlings at Sebeta, central Ethiopia, was
foodborne enteritis outbreaks have been solely or par- reported [79]. Vibrio spp., Salmonella, Shigella and E. coli
tially attributable to P. shigelloides [71]. Another species were detected from surface water and sediment samples
that is persistently detected in freshwater environments of Lake Ziway and drinking water system of Batu (former
and stands among major causes of food-/waterborne Ziway) town, Ethiopia [80].
human illnesses [72], but which could not be detected in From the total 410 fish samples examined, six were
the water samples of the current Lakes was S. typhi. found contaminated with Shiga toxin-producing E. coli
Detection of the common fecal coliforms (E. coli, K. strain in Ethiopia [81]. The isolates were resistant to
pneumonia, E. aerogenes) and Salmonella spp. in all of ampicillin and streptomycin disks. However, ciprofloxa-
the sample types especially in processed fish, signals the cin, gentamicin and nalidixic acid were found effective
danger of passage of these pathogens and their toxins to in inhibiting the growth of all of the isolates. Vibrio,
man via infected and contaminated fish products. Sal- Escherichia, Aeromonas, Pseudomonas, Salmonella and
monella spp. and fecal coliforms were detected in 42% of Streptococcus were detected from Nile tilapia in Hawassa
water samples and 64% of processed fish samples in this with the bacterial population significantly higher in the
study. Shigella spp. and Salmonella spp. are pathogenic intestine than in the liver [82]. A short review on bacte-
bacteria found in animal or human reservoir and con- rial pathogens of fish presented pathogenic and zoonotic
tamination of fish products by these bacteria is almost bacteria such as Edwardsiella, Salmonella, Escherichia,
always due to poor hygiene. Staphylococcus, Vibrio and Aeromonas recovered from
Except A. sobria, all the others species were detected fish from various parts of Ethiopia [83]. A more recent
from tilapia in different frequencies. Similarly, all bacteria molecular study that analyzed the diversity of microbiota
species were also detected from catfish with the excep- in different sections of tilapia gut found more diversity in
tion of P. shigelloides. However, C. freundii, C, koseri and Lake Chamo than Lake Hawassa [40].
V. parahemolyticus were not isolated from carps. Even In some countries like Poland [84] and Malaysia [85],
though it seemed that some bacteria species tended to the emergence of hitherto unreported pathogenic fish
be specifically associated with a particular fish species, bacteria is becoming evident. Thus, fish bacteria detec-
the association was not statistically significant. Different tion methods in Ethiopia must take into account less
studies reported the occurrence and antimicrobial resist- known and unreported ones as well. Moreover, exploring
ance of A. sobria, E. tarda, P. shigelloides, P. aeruginosa, possible reciprocal transmission of potential pathogenic
Citrobacter spp. and Klebsiella spp. from tilapia and cat- bacteria from wild fish to aquaculture, and domestic ani-
fish [73–76]. mals or humans is essential. This will contribute towards
During the study period, differences were observed in microbial-source-detection investigations.
the bacterial prevalence and frequency across the sam- This work will serve as an initial step to establish a base-
pling sites of each Lake. This may be attributed to the line dataset of microbial communities associated with
relative distance and degree of exposure to a nearby pol- wild freshwater fish in Ethiopia. But, it has certain nota-
lution source around. ble limitations. It neither quantified the detected bacte-
Although the report on fish bacteria and their occur- ria, nor molecularly identified them, and no antibiotic
rence in humans is limited in Ethiopia, there were some susceptibility test was done. The results would have been
efforts. Among the bacteria found in this study, E. coli, more robust if samples from fish skin and gills which are
Klebsiella spp., Enterobacter spp., Citrobacter spp., and gateway routes of transient or resident microbiota and/or
Aeromonas spp. which are enterotoxin-producing were potential pathogenic bacteria have been included. More-
detected in stools of Ethiopian children with diarrhoeal over, the study did not assess seasonal patterns of both
disease in the late 1970s [77]. E. tarda was isolated from water quality and fish microbiota.
the liver of a tilapia from Lake Ziway for the first time for
the Lake [15]. The other bacteria detected in this same Conclusion
study were E. coli, Kebsiella oxyloca, Citrobacter spp. and Despite the fact that the physicochemical parameters of
Yersinia enterocolitica. Another investigator [78] isolated the water samples were within normal range at which
Aeromonas spp. including A. sobria, E. tarda, Vibrio spp., most freshwater fish are non-stressed; the 20% prevalence
E. aerogenes P. shigelloides, E. coli, K. pneumoniae, Shi- of clinically sick fish is a source of concern. Moreover, the
gella spp., Citrobacter spp. from fish of Lake Tana. The bacteria identified from water, live fish and processed fish
author also reported that all the bacterial species, which samples are potential pathogens of fish and man, spoilage
Dissasa et al. BMC Veterinary Research (2022) 18:439 Page 12 of 14

agents and indicators of environmental contamination. Author details

1
Institute of Biotechnology, Addis Ababa University, Addis Ababa, Ethiopia.
In processed fish particularly, hygiene indicator bacteria 2
Department of Zoological Sciences, College of Natural and Computational
occurred at higher level suggesting that fresh processed Sciences, Addis Ababa University, Addis Ababa, Ethiopia. 3 Department
fish available in fish markets of Batu town likely act as a of Microbial, Cellular and Molecular Biology, College of Natural and Computa-
tional Sciences, Addis Ababa University, Addis Ababa, Ethiopia.
reservoir of pathogenic bacteria. Moreover, the identified
bacteria species are zoonotic and associated with food- Received: 30 December 2021 Accepted: 7 November 2022
borne illnesses. This has implications for the fish market
and consumer health. Therefore, consistent adherence to
simple hygienic steps is advisable. Further, establishing a
practice of regular inspection of fish products and source References
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