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.

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.

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.