Antimicrobial Resistance Profiles of E. coli Isolated From Pooled Samples of Sick, Farm, and Market Chickens in Nairobi County, Kenya.

Bacterial antimicrobial resistance (AMR) is a global threat to both human and animal health. This is mainly because the same antimicrobial molecules are used for the treatment and prophylaxis of bacterial diseases in both cases, and about 60% of human pathogens are shared with animals. For effective control of AMR in any country, the current situation has to be established; this is done through surveillance exercises. In Kenya, there is scanty data on the prevailing AMR situation, especially in animals. This paper reports on AMR profiles of 54 E. coli strains isolated from chickens in a cross-sectional study, out of which 36/54 (72%) were from clinically ill chickens, 11/54 (22%) were from farm chickens, and 7/54 (9.7%) were from slaughtered chicken, respectively. All 54 isolates exhibited varying antimicrobial resistance profiles with the majority showing resistance to Ampicillin (85.22%), Tetracycline (66.7%), Co-trimoxazole (57.4%), and Streptomycin (40.7%). Very few isolates were resistant to Amoxicillin and Gentamicin (each at 3.7%), Ampicillin (11.1%), and Nalidixic acid (24.1%). A total of 44/54 (81.5%) showed multiple resistance to up to 6 antimicrobial agents. This information will augment current data on the AMR status of bacteria harbored by chickens in Kenya. It will also inform policymakers in their fight against AMR.

Molecular Detection of Virulence-Associated Markers in Campylobacter coli and Campylobacter jejuni Isolates From Water, Cattle, and Chicken Faecal Samples From Kajiado County, Kenya.

Campylobacter is a zoonotic foodborne pathogen that is often linked with gastroenteritis and other extraintestinal infections in humans. This study is aimed at determining the genetic determinants of virulence-encoding genes responsible for flagellin motility protein A (flaA), Campylobacter adhesion to fibronectin F (cadF), Campylobacter invasion antigen B (ciaB) and cytolethal distending toxin (cdt) A (cdtA) in Campylobacter species. A total of 29 Campylobacter coli isolates (16 from cattle, 9 from chicken, and 4 from water samples) and 74 Campylobacter jejuni isolates (38 from cattle, 30 from chicken, and 6 from water samples) described in an earlier study in Kajiado County, Kenya, were examined for the occurrence of virulence-associated genes using polymerase chain reaction (PCR) and amplicon sequencing. The correlations among virulence genes were analyzed using Pearson's correlation coefficient (R) method. Among the 103 Campylobacter strains screened, 89 were found to harbour a single or multiple virulence gene(s), giving an overall prevalence of 86.4%. C. jejuni strains had the highest prevalence of multivirulence at 64.9% (48/74), compared to C. coli (58.6%, 17/29). The ciaB and flaA genes were the most common virulence genes detected in C. jejuni (81.1% [60/74] and 62.2% [46/74], respectively) and in C. coli (each at 62.1%; 18/29). Campylobacter isolates from chicken harboured the most virulence-encoding genes. C. jejuni strains from chicken and cattle harboured the highest proportions of the cdtA and ciaB genes, respectively. All the C. coli strains from water samples harboured the cadF and flaA genes. The results obtained further revealed a significant positive correlation between cadF and flaA (R = 0.733). C. jejuni and C. coli strains from cattle, chicken, and water harbour virulence markers responsible for motility/colonization, invasion, adherence, and toxin production, evoking their important role in campylobacteriosis development among humans and livestock. The identification of cattle, chicken, and water samples as reservoirs of virulent Campylobacter spp. highlights the possible risk to human health. These data on some virulence genes of Campylobacter will assist food safety and public health officials in formulating policy statements.

Prevalence and risk factors associated with the occurrence of Campylobacter sp. in children aged 6-24 months in peri-urban Nairobi, Kenya.

Introduction: Campylobacter bacteria is a major cause of foodborne-related bacterial gastroenteritis in humans worldwide. It is known to cause diarrhea in young children which has been shown to directly affect their weight and height as a result of malnutrition. Severe cases of diarrhea can also lead to death. Most of the burden is experienced in resource-limited countries in Africa and Southeast Asia where the disease is linked to poor hygiene and sanitation. The objective of this study was to determine the prevalence of Campylobacter in children aged between 6 and 24 months in Nairobi, Kenya and identify potential risk factors associated with their occurrence. Methods: A cross-sectional study was carried out between May to December 2021. A total of 585 randomly selected households were visited in two wards (Uthiru/Ruthimitu and Riruta) in Dagoretti South sub-county, Nairobi. A questionnaire regarding how children's food is handled, the major foods consumed, sanitation and hygiene, and animal ownership was conducted among caregivers to identify associated risk factors. Stool samples were collected from 540/585 children and screened for the presence of Campylobacter using culture-based methods and confirmed through PCR. Results: Of the 540 children's stool samples processed, Campylobacter isolates were detected in 4.8% (26/540). Campylobacter jejuni (C. jejuni) was the most common species in 80.8% of positive samples compared to Campylobacter coli (C. coli) in 26.9% of samples. In six samples, both C. jejuni and C. coli were isolated, while in four samples, it was not possible to speciate the Campylobacter. Drinking cow's milk (OR 4.2, 95% CI 1.4 - 12.6) and the presence of animal feces in the compound (OR 3.4, 95% CI 1.1 - 10.3) were found to be statistically associated with Campylobacter carriage in children. Discussion: The carriage of Campylobacter in children in this community indicates a need for further investigation on source attribution to understand transmission dynamics and inform where to target interventions. Awareness creation among caregivers on good personal and food hygiene is needed, including boiling milk before consumption. Implementation of biosecurity measures at the household level is highly recommended to reduce contact between animals and humans.

Seroprevalence of Peste des Petits Ruminants and Contagious Caprine Pleuropneumonia Coinfections in Goats in Kwale County, Kenya.

Goats are among the most important small ruminants affected by Peste des Petits ruminants (PPR) and contagious caprine pleuropneumonia (CCPP) diseases, two of the most significant constraints worldwide to the production of small ruminant species. Herein, the competitive enzyme-linked immunosorbent assay (cELISA) and the latex agglutination test (LAT) were used to determine the coinfections of PPR and CCPP in goats in Kwale County on Kenya's South Coast. A total of 368 serum samples were collected from goats of various ages and sexes exhibiting respiratory distress in the four subcounties of Kwale County (Kinango, Lunga Lunga, Matuga, and Msambweni) and screened for PPR and CCPP antibodies. Of the 368 goats sampled, 259 (70.4%) were females and 109 (29.6%) were males, and 126 (34.2%), 71 (19.3%), 108 (29.3%), and 63 (17.1%) samples were collected from Kinango, Matuga, Lunga Lunga, and Msambweni, respectively. The overall PPR seropositivity rate was 48.6% (179/368); rates in Kinango, Lunga Lunga, Matuga, and Msambweni were 70.6%, 29.6%, 49.3%, and 36.5%, respectively. The overall CCPP seropositivity rate was 45.4% (167/368), while rates in Kinango, Lunga Lunga, Matuga, and Msambweni were 51.6%, 49.1%, 36.6%, and 36.5%, respectively. Notably, the seropositivity of PPR was higher in male (53.3%) than in female (46.72%) goats, though not statistically significant. In addition, the CCPP seropositivity rates were not significantly different between male (44.0%) and female (45.9%) goats. Regarding age, the PPR seropositivity rates were 45.9%, 55.8%, and 52.3% in adults, kids, and weaners, respectively. For CCPP, the seropositivity rates were 48.3%, 40.4%, and 42.3% in adults, kids, and weaners, respectively. The coinfection rate of PPR and CCPP was 22.3% (82/368). Despite the high coinfection, univariate analysis revealed no relationship between PPR and CCPP infections. However, given the high PPR and CCPP infection rates, as a result of separate or coinfection, there is a need to upscale or intensify vaccination in the county.

Antimicrobial Usage, Susceptibility Profiles, and Resistance Genes in Campylobacter Isolated from Cattle, Chicken, and Water Samples in Kajiado County, Kenya.

Campylobacter organisms are the major cause of bacterial gastroenteritis and diarrhoeal illness in man and livestock. Campylobacter is growingly becoming resistant to critically crucial antibiotics; thereby presenting public health challenge. This study aimed at establishing antimicrobial use, susceptibility profiles, and resistance genes in Campylobacter isolates recovered from chicken, cattle, and cattle-trough water samples. The study was conducted between October 2020 and May 2022 and involved the revival of cryopreserved Campylobacter isolates confirmed by PCR from a previous prevalence study in Kajiado County, Kenya. Data on antimicrobial use and animal health-seeking behaviour among livestock owners (from the same farms where sampling was done for the prevalence study) were collected through interview using a pretested semistructured questionnaire. One hundred and three isolates (29 C. coli (16 cattle isolates, 9 chicken isolates, and 4 water isolates) and 74 C. jejuni (38 cattle isolates, 30 chicken isolates, and 6 water isolates)) were assayed for phenotypic antibiotic susceptibility profile using the Kirby-Bauer disk diffusion method for ampicillin (AX), tetracycline (TE), gentamicin (GEN), erythromycin (E), ciprofloxacin (CIP), and nalidixic acid (NA). Furthermore, detection of genes conferring resistance to tetracyclines (tet (O), ß-lactams (bla OXA-61), aminoglycosides (aph-3-1), (fluoro)quinolones (gyrA), and multidrug efflux pump (cmeB) encoding resistance to multiple antibiotics was detected by mPCR and confirmed by DNA sequencing. The correlation between antibiotic use and resistance phenotypes was determined using the Pearson's correlation coefficient (r) method. Tetracyclines, aminoglycosides, and ß-lactam-based antibiotics were the most commonly used antimicrobials; with most farms generally reported using antimicrobials in chicken production systems than in cattle. The highest resistance amongst isolates was recorded in ampicillin (100%), followed by tetracycline (97.1%), erythromycin (75.7%), and ciprofloxacin (63.1%). Multidrug resistance (MDR) profile was observed in 99 of 103 (96.1%) isolates; with all the Campylobacter coli isolates displaying MDR. All chicken isolates (39/39, 100%) exhibited multidrug resistance. The AX-TE-E-CIP was the most common MDR pattern at 29.1%. The antibiotic resistance genes were detected as follows: tet (O), gyrA, cmeB, bla OXA-61 , and aph-3-1 genes were detected at 93.2%, 61.2%, 54.4%, 36.9%, and 22.3% of all Campylobacter isolates, respectively. The highest correlations were found between tet (O) and tetracycline-resistant phenotypes for C. coli (96.4%) and C. jejuni (95.8%). A moderate level of concordance was observed between the Kirby-Bauer disk diffusion method (phenotypic assay) and PCR (genotypic assay) for tetracycline in both C. coli (kappa coefficient = 0.65) and C. jejuni (kappa coefficient = 0.55). The study discloses relatively high resistance profiles and multidrug resistance to antibiotics of critical importance in humans. The evolution of the multidrug-resistantCampylobacter isolates has been linked to the use and misuse of antimicrobials. This poses a potential hazard to public and animal health, necessitating need to reduce the use of antibiotics in livestock husbandry practice coupled with stringent biosecurity measures to mitigate antimicrobial resistance.

Frequency and diversity of carbapenemase-producing Enterobacterales recovered from untreated wastewater impacted by selective media containing cefotaxime and meropenem in Ohio, USA.

As safe agents of last resort, carbapenems are reserved for the treatment of infections caused by multidrug-resistant organisms. The impact of ß-lactam antibiotics, cefotaxime, and meropenem on the frequency and diversity of carbapenemase-producing organisms recovered from environmental samples has not been fully established. Therefore, this methodological study aimed at determining ß-lactam drugs used in selective enrichment and their impact on the recovery of carbapenemase-producing Enterobacterales (CPE) from untreated wastewater. We used a longitudinal study design where 1L wastewater samples were collected weekly from wastewater treatment plant (WWTP) influent and quarterly from contributing sanitary sewers in Columbus, Ohio USA with 52 total samples collected. Aliquots of 500 mL were passed through membrane filters of decreasing pore sizes to enable all the water to pass through and capture bacteria. For each sample, the resulting filters were placed into two modified MacConkey (MAC) broths, one supplemented with 0.5 µg/mL of meropenem and 70 µg/mL of ZnSO4 and the other supplemented with 2 µg/mL cefotaxime. The inoculated broth was then incubated at 37° C overnight, after which they were streaked onto two types of correspondingly-modified MAC agar plates supplemented with 0.5 µg/mL and 1.0 µg/mL of meropenem and 70 µg/mL of ZnSO4 and incubated at 37°C overnight. The isolates were identified based on morphological and biochemical characteristics. Then, up to four distinct colonies of each isolate's pure culture per sample were tested for carbapenemase production using the Carba-NP test. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) MALDI-TOF MS was used to identify carbapenemase-producing organisms. In total 391 Carba-NP positive isolates were recovered from the 52 wastewater samples: 305 (78%) isolates had blaKPC, 73 (19%) carried blaNDM, and 14 (4%) harbored both blaKPC and blaNDM resistance genes. CPE genes of both blaKPC and blaNDM were recovered in both types of modified MAC broths, with 84 (21%) having a blaKPC gene, 22 (6%) carrying blaNDM and 9 (2%) harbored both a blaKPC and blaNDM of isolates recovered from MAC medium incorporated with 0.5ug/mL meropenem and 70ug/mL ZnSO4. The most prevalent isolates were Klebsiella pneumoniae, Escherichia coli, and Citrobacter spp.

Human brucellosis in Baringo County, Kenya: Evaluating the diagnostic kits used and identifying infecting Brucella species.

Human brucellosis diagnosis has been a challenge in Brucella-endemic areas. In Kenya, diagnosis is usually carried out using Febrile Brucella Antigen agglutination test (FBAT) whose performance is not well documented. This paper reports on the sensitivity and specificity of the FBAT used for brucellosis diagnosis on blood samples/serum collected in three healthcare facilities in Baringo County, Kenya, and on Brucella species present in the study area. The FBAT test results at the hospitals were used to guide patient management. Patients who visited the hospital's laboratory with a clinician's request for brucellosis testing also filled a questionnaire to assess knowledge and attitudes associated with transmission of the disease in the study area. The remaining serum samples were tested again using FBAT and Rose Bengal Plate Test (RBPT) within a month of blood collection at the University Nairobi Laboratory. The two rapid tests were then compared, with respect to brucellosis diagnostic sensitivity and specificity. To identify infecting Brucella species, a proportion 43% (71/166) of the blood clots were analyzed by multiplex polymerase chain reaction (PCR) using specific primers for B. abortus, B. melitensis, B. ovis and B. suis. Out of 166 serum samples tested, 26.5% (44/166) were positive using FBAT and 10.2% (17/166) positive using RBPT. The sensitivity and specificity of FBAT compared to RBPT was 76.47% and 71.19%, respectively while the positive and negative predictive values were 29.55% and 96.72%, respectively. The FBAT showed higher positivity then RBPT. The difference in sensitivity and specificity of FBAT and RBPTs was relatively low. The high FBAT positivity rate would be indication of misdiagnosis; this would lead to incorrect treatment. Brucella abortus was detected from 9.9% (7/71) of the blood clots tested; no other Brucella species were detected. Thus human brucellosis, in Baringo was mainly caused by B. abortus.

Seroprevalence and spatial distribution of livestock brucellosis using three serological tests in Kajiado County, Kenya.

Background: Brucellosis is a serious zoonotic infection with a global socioeconomic impact on both the livestock industry and human health. In Kenya, brucellosis is endemic but there is limited information on the true burden of the disease due to weak or peace-meal surveillance. The true burden and spread of animal brucellosis in Kajiado County is not known. Aim: The aim of the study was to determine the current seroprevalence and spatial distribution of livestock brucellosis in Kajiado County and also to compare the three serological tests, namely; Rose Bengal plate test (RBPT), indirect enzyme-linked immunosorbent assay (i-ELISA), and competitive ELISA (c-ELISA) in the detection of seropositive animals. Methods: A cross-sectional study was undertaken in 5 sub-counties and 13 wards, where a total of 782 serum samples from unvaccinated bovine (n = 278; 34 herds), ovine (n = 256; 25 flocks), and caprine (n = 248; 28 flocks), were screened for anti-Brucella antibodies using RBPT, i-ELISA, and c-ELISA tests, in parallel. Results: Overall animal seroprevalence was 6.91% (54/782); while that for bovine, ovine, and caprine was 18.35% (51/278), 0.78% (2/256), and 0.4% (1/248), respectively. Bovine seroprevalence was 2.2% (6/278), 14.4% (40/278), and 4.7% (13/278) in RBPT, i-ELISA, and c-ELISA tests, respectively; while ovine 0.78% (2/256) and caprine 0.4% (1/248) were positive only in c-ELISA. Bovine herd seropositivity was 67.65% (23/34), whereas ovine and caprine flock seropositivities were 8% (2/25) and 3.6% (1/28), respectively. Conclusion: The findings indicate a moderate seroprevalence of brucellosis in bovine, while that of ovine and caprine was low in Kajiado County. Indirect ELISA was found superior to both c-ELISA and RBPT in detecting bovine seropositive animals, while c-ELISA was superior to both RBPT and i-ELISA in detecting seropositive ovines and caprines. These results will contribute to baseline data for further study of Brucella infection and a starting point for the formulation of a strategy for the control of brucellosis in Kajiado County.

Characteristics of Staphylococcus aureus Isolated from Patients in Busia County Referral Hospital, Kenya.

Staphylococcus aureus is an important pathogen associated with hospital, community, and livestock-acquired infections, with the ability to develop resistance to antibiotics. Nasal carriage by hospital inpatients is a risk for opportunistic infections. Antibiotic susceptibility patterns, virulence genes and genetic population structure of S. aureus nasal isolates, from inpatients at Busia County Referral Hospital (BCRH) were analyzed. A total of 263 inpatients were randomly sampled, from May to July 2015. The majority of inpatients (85.9%) were treated empirically with antimicrobials, including ceftriaxone (65.8%) and metronidazole (49.8%). Thirty S. aureus isolates were cultured from 29 inpatients with a prevalence of 11% (10.3% methicillin-susceptible S. aureus (MSSA), 0.8% methicillin resistant S. aureus (MRSA)). Phenotypic and genotypic resistance was highest to penicillin-G (96.8%), trimethoprim (73.3%), and tetracycline (13.3%) with 20% of isolates classified as multidrug resistant. Virulence genes, Panton-Valentine leukocidin (pvl), toxic shock syndrome toxin-1 (tsst-1), and sasX gene were detected in 16.7%, 23.3% and 3.3% of isolates. Phylogenetic analysis showed 4 predominant clonal complexes CC152, CC8, CC80, and CC508. This study has identified that inpatients of BCRH were carriers of S. aureus harbouring virulence genes and resistance to a range of antibiotics. This may indicate a public health risk to other patients and the community.

Multi-Drug Resistant Staphylococcus aureus Carriage in Abattoir Workers in Busia, Kenya.

Abattoir workers have been identified as high-risk for livestock-associated Staphylococcus aureus carriage. This study investigated S. aureus carriage in abattoir workers in Western Kenya. Nasal swabs were collected once from participants between February-November 2012. S. aureus was isolated using bacterial culture and antibiotic susceptibility testing performed using the VITEK 2 instrument and disc diffusion methods. Isolates underwent whole genome sequencing and Multi Locus Sequence Types were derived from these data. S. aureus (n = 126) was isolated from 118/737 (16.0%) participants. Carriage was higher in HIV-positive (24/89, 27.0%) than HIV−negative participants (94/648, 14.5%; p = 0.003). There were 23 sequence types (STs) identified, and half of the isolates were ST152 (34.1%) or ST8 (15.1%). Many isolates carried the Panton-Valentine leucocidin toxin gene (42.9%). Only three isolates were methicillin resistant S. aureus (MRSA) (3/126, 2.4%) and the prevalence of MRSA carriage was 0.4% (3/737). All MRSA were ST88. Isolates from HIV-positive participants (37.0%) were more frequently resistant to sulfamethoxazole/trimethoprim compared to isolates from HIV-negative participants (6.1%; p < 0.001). Similarly, trimethoprim resistance genes were more frequently detected in isolates from HIV-positive (81.5%) compared to HIV-negative participants (60.6%; p = 0.044). S. aureus in abattoir workers were representative of major sequence types in Africa, with a high proportion being toxigenic isolates. HIV-positive individuals were more frequently colonized by antimicrobial resistant S. aureus which may be explained by prophylactic antimicrobial use.

Seasonal Prevalence and Molecular Identification of Thermophilic Campylobacter from Chicken, Cattle, and Respective Drinking Water in Kajiado County, Kenya.

Thermophilic Campylobacter species are a leading cause of human gastroenteritis throughout the world and have been implicated in reproductive disorders (abortion), mastitis, enteritis, and/or diarrhoea in livestock. A cross-sectional survey was conducted in Kajiado County to determine prevalence, seasonality, and molecular detection of thermophilic Campylobacter species (with emphasis on C. jejuni, C. coli, and other thermophilic Campylobacter species) in chicken, cattle, and respective pooled drinking water. A total of 457 samples comprising 265 cattle rectal swabs, 142 chicken cloacal swabs, and 50 trough water samples were collected from 55 randomly selected smallholder farms. Individual samples were subjected to standard techniques for isolation and biochemical tests, followed by singleplex polymerase chain reaction (sPCR) assays for identification and confirmation of genus and species. Overall, thermophilic Campylobacter prevalence was 35.4% (95% confidence interval (95% CI) = 31.0-39.8), with C. jejuni dominating at 55.6% (95% CI = 47.9-63.3%) over C. coli in all sample types. The highest thermophilic Campylobacter prevalence was observed in cloacal swabs of live chicken at 44.4% (95% CI = 36.2-52.6%), followed by rectal swabs from live cattle at 30.9% (95% CI = 25.3-36.5%). Water samples from cattle drinkers/trough were found to be contaminated at 34% (95% CI = 20.9-47.1%). The isolation rate was higher in cattle under the confinement system (44.3%) (95% CI = 36.1-52.5%) than in those under the free-roaming grazing system. Thermophilic Campylobacter species were isolated in both seasons, with higher prevalence (39.8% (95% CI = 33.6-45.9)) recorded during rainy and cold season in all sample types except for water. There was significant (P < 0.05) association between season and thermophilic Campylobacter occurrence, even though there were no statistical differences in the prevalence values across the two seasons. Results of this study demonstrate that cattle, chicken, and respective drinking water harbour potentially pathogenic thermophilic campylobacters, with C. jejuni being widely distributed among farms. It is possible that seasonal variations and cattle confinement result in differences in thermophilic Campylobacter carriage. Further epidemiological and phylogenetic studies comparing distribution of thermophilic Campylobacter spp. isolates in livestock, environmental, and human samples are recommended to establish source attribution to reduce the impact of resultant diseases for the wellbeing of public and livestock.

Village-Indigenous Chicken Bacterial Carriage after the Heavy Rains of 2018, Kenya: Indicator on Environmental Contamination with Pathogenic/Zoonotic Bacteria.

Food borne diseases are one of the major human disease conditions worldwide. Most of them are of bacterial origin and chickens are a major source of such bacteria; they are consumed at high rate worldwide and tend to harbor the zoonotic bacteria without showing signs of illness. Running rain water tends to increase environmental contamination, since it carries various substances from one area to another; this results in village-indigenous chickens picking more bacteria from the environment as they roam/scavenge around for food. Thus, after the rain, the chickens' intestinal contents may contain more bacteria quantity-wise and type-wise. This study was carried-out to determine whether that was the case after heavy rains of 2018.120 intestine samples were collected from indigenous chickens from three slaughterhouses in Nairobi for bacterial quantification using the Miles and Misra technique; bacterial isolation and identification were carried out using standard bacteriological procedures. Intestines from the slaughterhouses had different mean bacterial counts: Kangemi had the highest (1.3 × 1012 colony-forming units per ml), followed by Burma (5.6 × 1011), then Kariokor (4.7 × 1011). E. coli was the most isolated at 85.8%, followed by genera Staphylococcus (55%), Streptococcus (43.3%), Bacillus (41.66%), Listeria (38.3%), Proteus (24.16%), Klebsiella (7.5%), Campylobacter (2.5%), Pseudomonas (6%), and Streptobacillus (0.83%). The study showed that the indigenous chickens carry a variety of bacteria in types and numbers, some of them being zoonotic. Apart from picking more bacteria as a result of environmental contamination during rainy season, the weather and bird-handling further stress the birds, thus contributing to higher bacterial multiplication and higher bacterial carriage. If slaughter is not done right, these intestinal bacteria can easily cause contamination of respective chicken meat; thus, if pathogenic, it can cause food poisoning to consumers of the meat. Therefore, it is recommended that precaution be taken while slaughtering chickens for consumption. In addition, where possible, free-range indigenous chickens be confined during rainy seasons to reduce their exposure to contaminated environment.

Antibiotic and Disinfectant Susceptibility Patterns of Bacteria Isolated from Farmed Fish in Kirinyaga County, Kenya.

Fish bacterial pathogens cause diseases which result in a considerable economic impact on the aquaculture industry, necessitating the use of antimicrobials for their control. However, intensive and indiscriminate use of antimicrobials has led to increased occurrence of drug resistance in pathogenic bacteria, as well as normal flora. The aim of the current study was to determine the susceptibility patterns of bacteria isolated from fish, with respect to some commonly used antibiotics and disinfectants. Bacteria were isolated between December 2017 and April 2018 from farmed Nile tilapia, African catfish, goldfish, and koi carp in Kirinyaga County, Kenya. Antibiotic and disinfectant susceptibility patterns of 48 isolates belonging to the genera Aeromonas, Proteus, Klebsiella, Citrobacter, Salmonella, Streptococcus, Pseudomonas, Escherichia, Serratia, and Micrococcus were established using the Kirby-Bauer disc diffusion method and agar well diffusion technique, respectively. The antibiotics evaluated included ampicillin, tetracycline, co-trimoxazole, streptomycin, kanamycin, gentamicin, co-trimoxazole, and chloramphenicol, while the disinfectants tested were quaternary ammonium compound, formalin, hydrogen peroxide, sodium hypochlorite, and iodine. All the bacteria except Micrococcus, Escherichia, and Salmonella species showed multiple drug resistance patterns. Streptococcus showed resistance to six antibiotics, while Proteus, Pseudomonas, and Serratia were resistant to five antibiotics. The multiple antibiotic resistance index ranged from 0.1 to 0.8, with Streptococcus spp. having the highest score value. All the organisms were sensitive to gentamicin, while co-trimoxazole and ampicillin showed the highest resistance at 73% (n = 34) and 62% (n = 31), respectively. Most of the disinfectants showed antibacterial activity with varying magnitudes. The isolates were 100% sensitive to hydrogen peroxide and formalin, but were resistant to sodium hypochlorite at recommended user-dilution. The study has shown that some of the bacterial isolates were resistant to common antibiotics and disinfectants; thus, it is recommended to include an antibiogram whenever making any therapeutic decision. The resistant bacteria may transmit resistance genes to other fish bacteria and also to human bacteria, thus making it difficult to treat the resultant disease(s); thus, there is a possibility that these resistant bacteria may be transmitted to humans who consume or handle the carrier fish. It is, therefore, advisable that fish are cooked properly before consumption, so as to kill bacteria that may be present.

Natural Concurrent Infections with Black Spot Disease and Multiple Bacteriosis in Farmed Nile Tilapia in Central Kenya.

Nile tilapia (Oreochromis niloticus) is the most cultured and available fish for Kenyan consumers, and therefore, any tilapine disease deprives them the valuable source of protein. Nile tilapia farm was diagnosed with severe concurrent black spot disease and multiple bacteriosis using gross lesions and parasitological, histopathology, and standard bacteriological procedures. A total of 25 fish were sampled and inspected, and all of them had raised, macroscopic 1 mm-sized black spot lesions. The mean intensity of black spots per fish was 728 with an abundance of 2-1740 metacercariae cysts per fish. A high intensity of black spot infestation was observed in the fins (43.9%), skin and underlying muscles (18.3%), and gills (18%). In addition, histopathological data confirmed presence of a metacercaria of Neascus spp. as the aetiological agent of black spot disease. Furthermore, a thick fibrous capsule around the metacercaria, black pigment melanomacrophages, and moderate muscle atrophy were observed. The most prevalent bacteria isolated were Aeromonas, Enterobacter cloacae, Klebsiella pneumoniae, and Micrococcus luteus. Physicochemical parameters of pond water were temperature (28.2°C), dissolved oxygen (4.2 mgl-1), pH (8.5), ammonia free nitrogen (15.8 mgl-1), alkalinity (112 mgl-1), hardness (68 mgl-1), nitrites (0.058 mgl-1), nitrates (58 mgl-1), and phosphates (0.046 mgl-1). However, the levels of nitrates, nitrites, alkalinity, and ammonia free nitrogen exceeded the recommended limits. In conclusion, these findings suggest that coinfections by these organisms coupled by water quality-related stress can be associated with low-grade mortality observed in postfingerling tilapia as well as reduced growth. The authors recommended immediate destocking, thorough disinfection, and control of piscivorous birds. Moreover, attention ought to be geared towards prevention of parasitic infestations in fish so as to minimize fish deaths related to secondary bacteriosis. Further experimental studies should be carried out to elucidate the relationship of these pathogens.

Fish Husbandry Practices and Water Quality in Central Kenya: Potential Risk Factors for Fish Mortality and Infectious Diseases.

Fish mortality has an enormous impact on the aquaculture industry by reducing fish production and slowing industrial growth. A cross-sectional study was carried out in Kirinyaga County, Central Kenya, to evaluate potential risks of fish mortality and disease transmission and suitability of pond water for rearing fish. A semistructured questionnaire that focused on general information, management practices, and disease history was administered to 92 small-scale fish farmers. Parasitological examination of fish sampled from selected farms (farms that were reporting mortality at the time of sampling) was done by following the standard procedure. Water quality parameters for 33 ponds were evaluated in situ (recorded on pond site) and ex situ (analysed at the laboratory) following the standard methods. The risks were assessed by adjusted odds ratio based on univariate regression analysis. Prevalent fish husbandry practices that were found to be associated with fish mortality and acquisition of pathogens in the study area were the use of raw livestock manure (0R = 1.500), high fish stocking density (0R = 1.168), and feeding fish on homemade rations (0R = 1.128). Parasitological investigation found infestation with Diplostomum spp., Dactylogyrus spp., Clinostomum spp., and Piscicola leeches. Water temperature and pH were found fit for rearing fish. Of the 33 fishpond water samples tested, 1 (3%) and 6 (18%) exceeded the recommended limits of <100 mg/L and <0.2 mg/L of nitrate and nitrite, respectively. Of the 29 fishpond water tested, 15 (59%) exceeded the recommended limits of <100 mg/L of total ammonia. The findings show that the use of raw livestock manure, high fish stocking density, high nitrates and nitrites, and high ammonia levels in fishponds are potential risk factors for fish mortality and acquisition of infectious pathogens in a pond environment in a rural setup, in Central Kenya. There is a need to address the above factors in small-scale farming practices to minimize fish loss and also to prevent the occurrence and spread of infectious pathogens.

Haemato-biochemical changes and prevalence of parasitic infections of indigenous chicken sold in markets of Kiambu County, Kenya.

This study aimed at determining parasitic prevalence and probable haemato-biochemical changes that may occur from parasitic infections in marketed indigenous chickens in Kiambu County, Kenya. Thirty adult chickens were purchased and examined for ectoparasites, haemoparasites and haemato-biochemical changes. Post mortem was conducted to recover gastro-intestinal parasites and fecal samples taken for egg/oocyst counts. Forty-seven percent (14/30) of chickens examined were in poor body condition, 43% (13/30) in fair and 10% (3/30) in good body condition. Ectoparasites infection prevalence was 66.7% (20/30). Four haemoparasites were isolated. Overall helminths prevalence was 86.6% (26/30), nematodes at 76.7% (23/30) and cestodes at 40% (12/30). After processing fecal samples, 30% (9/30) were positive for helminth eggs and 30% (9/30) had coccidial oocysts. Relative to normal values, total erythrocyte count was low and total leucocyte count with band cells high. Mean haematocrit and heterophil values were high (p=0.0005; p=0.0061). Mean lymphocyte count was low (p=0.0128) in chickens with ectoparasitic infestation. Eosinophils increased significantly (p=0.0363) although mean erythrocytes counts decreased (p=0.0176), in chickens with gastrointestinal parasites. Creatine phosphokinase and blood glucose levels were high, serum protein and albumin levels were low. Blood glucose level decreased significantly (p=0.0239) and total plasma protein increased (p=0.045) in chickens with Haemoproteus spp. infection. The study showed, ecto- and endo-parasites are prevalent and may contribute to alteration of haemato-biochemical parameters of sub-clinically infected marketed indigenous chickens. These results are expected to contribute towards and encourage usage of clinico-pathological parameter testing as a measure of poultry health status for enhanced poultry disease diagnoses.

Hematologic Values of Healthy and Sick Free-ranging Lesser Flamingos ( Phoeniconaias minor) in Kenya.

We determined hematologic parameters of five healthy and nine sick free-ranging Lesser Flamingos ( Phoeniconaias minor) from Lake Nakuru, Kenya. Heterophilia and lymphopenia were evident in sick birds, with up to 7.5-fold higher heterophil-to-lymphocyte ratio in sick birds compared to healthy birds. Leucopenia was present in a few sick birds. A higher than normal packed cell volume was observed in birds that had evidence of acute disease, whereas a lower than normal packed cell volume was seen in birds with evidence of prolonged sickness. Healthy birds had higher total white blood cell counts and lymphocyte counts and lower heterophil counts than zoo flamingos. Most sick birds were diagnosed with septicemia, occasionally with fibrinous exudation into the coelomic cavities. One bird had mycobacterial granulomas, one had a corynebacterium-associated wing abscess, and one had a wing fracture. We provide hematologic data for free-ranging Lesser Flamingos and compare the parameters of sick and healthy birds.

Risk factors associated with infectious bursal disease vaccination failures in broiler farms in Kenya.

Immunization together with application of biosecurity measures are the principal methods of preventing infectious bursal disease outbreaks in high-risk areas. However, outbreaks in vaccinated chicken flocks have been reported in many parts of the world as a result of factors of vaccine virus, animal, or vaccine handler. In Kenya, such outbreaks have been reported, but the causes have not been studied. This study aimed at determining the risk factors associated with vaccine handling leading to vaccine failure in broiler flocks in Kwale County, Kenya. Structured questionnaires and visual observations were used to collect data from 83 broiler farms, 6 breeding farms, and 17 vaccine outlets. Relative risk (RR) analysis was used to determine the association between identified potential risk factors and vaccination failure. Results show that vaccines were properly handled in all vaccine outlet shops. Breeding farms maintained high levels of biosecurity and employed standard vaccine handling practices. Basic biosecurity practices were poor in broiler farms. Broiler farms failed to meet all the recommended standard procedures for vaccine storage, reconstitution, and administration. Risk factors included poor vaccine storage (RR = 8.7) and use of few drinkers to administer vaccine (RR = 5.8); traces of disinfectants in drinkers used to administer live vaccine (RR = 2.8); use of wrong vaccine-infectious bronchitis instead of infectious bursal disease vaccine (RR = 2.1); and use of improper diluents (RR = 1.6). Broiler farmers need training on basic farm biosecurity measures and standard vaccine handling practices.

Effect of immunosuppression on newcastle disease virus persistence in ducks with different immune status.

This study was carried out to verify the possibility that ducks are sources of Newcastle disease (ND) virus infection for chickens in mixed flocks. Immunosuppressed (IS) and non immunosuppressed (NIS) birds, at three different antibody levels (medium, low and absent) were used; the titres having been induced through vaccination, and Immunosuppression done using dexamethazone. Each of the 3 respective groups was further divided into 2 groups of about 12 ducks each: one challenged with velogenic ND virus; the other not challenged. Selected ducks from all groups had their antibody titres monitored serially using hemagglutination inhibition test, while two birds from each of the challenged groups were killed and respective tissues processed for ND viral recovery, using chicken embryo fibroblasts. In general, antibody titres of IS and NIS challenged ducks were significantly higher than their unchallenged counterparts (P < 0.05). Non-challenged pre-immunised ducks had a progressive decrease in antibody levels; non-immunised ducks did not seroconvert. Newcastle disease virus was isolated from livers and kidneys of the challenged ducks throughout the experimental period; indicating a possibility of viral excretion, especially when the birds are stressed. It, therefore, provides another possible model of viral circulation within mixed flocks.

Viral nucleoprotein localization and lesions of Newcastle disease in tissues of indigenous ducks.

Localization of Newcastle disease viral nucleoprotein and pathological lesions was evaluated in tissues of 55 indigenous ducks (45 experimentally infected and 10 sentinel ones). In addition, ten Newcastle disease infected chickens were used to ensure that the virus inoculum administered to the ducks produced the disease in chickens, the susceptible hosts. Ducks were killed on day 1, 4, 8 and 14 post-infection. Post-mortem examination was done with six tissues (liver, spleen, lung, caecal tonsils, kidneys and brain) being collected from each bird. The tissues were preserved in 10% neutral formalin for 24 h. They were then transferred to 70% ethanol for histology and immunohistochemical staining. Airsacculitis, necrotic splenic foci, congested intestines, lymphoid depleted caecal tonsils and focal infiltrations by mononuclear cells were the main pathological lesions in infected ducks. Over 28.9% of the infected ducks had Newcastle disease viral nucleoprotein in macrophage-like large mononuclear cells in the caecal tonsils and kidney tubular epithelium. The viral antigens were located in the cytoplasm and nucleolus of the cells. The other organs had no detectable viral antigens. This study shows that the kidneys and caecal tonsils are the likely predilection sites for the virus in ducks. They thus need to be considered as diagnostic indicators for the viral carriage in ducks.