Isolation of vancomycin-resistant Enterococcus from apparently healthy human animal attendants, cattle and cattle wastes in Tanzania.

AIM: The study aimed to isolate and characterize Enterococcus species from apparently healthy waste attendants, cattle and cattle waste in Tanzania. Emphasis was given to antimicrobial resistance and in particular occurrence of vancomycin (VA)-resistant enterococci. METHODS AND RESULTS: Faecal samples were collected from healthy cattle, cattle waste attendants and cattle house wastes, and isolation of Enterococcus species was performed using Slanetz Bartley agar. Isolates were characterized with regard to species, antimicrobial susceptibility and presence of VA resistance genes. Enterococcus faecalis was the most prevalent species from all sources of isolation (43·5%), followed by Enterococcus faecium (38·4%). Isolates of E. faecium showed a higher number of phenotypic antimicrobial resistance than isolates of E. faecalis. Fifty-eight isolates, which showed resistance or intermediate resistance to VA by disc diffusion test, were analysed for VA-resistant Enterococcus (VRE) by PCR. The vanA gene was detected in 14 isolates of E. faecium and 12 isolates of E. faecalis, while vanB was detected in three isolates. No isolates were found to carry vanC1-gene. CONCLUSION: VRE was detected in both human and cattle samples, despite no known use of antimicrobial agents that can select for VRE in livestock in Tanzania. Enterococcus faecalis was the most commonly isolated species from cattle and humans. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides information on the prevalence of VRE in human and nonhuman samples in Tanzania calling for further studies on the origin of VRE in such isolates, since no selection mechanism in Tanzania are known.

Antimicrobial resistance in faecal samples from buffalo, wildebeest and zebra grazing together with and without cattle in Tanzania.

AIM: The aim of this study was to determine whether the practice of co-grazing with cattle and wild life constitutes a risk of transmission of antibiotic resistant bacteria to wild ungulates. METHODS AND RESULTS: Faecal samples were collected from buffalo (n = 35), wildebeest (n = 40), zebra (n = 40) and cattle (N = 20) from Mikumi National Park, Tanzania (MNP), where cattle is prohibited and from Ngorongoro Conservation Area (NCA) where co-grazing is practiced. The number of coliforms and enterococci resistant to selected antibiotics was determined. Wild life generally harboured higher number of resistant Escherichia coli and Enterococci than cattle, but with no general influence in wild life of co-grazing with cattle. Vancomycin-resistant Enterococci were detected in wild life samples, and E. coli resistant to cefotaxime and enrofloxacin were observed among isolates from all wild life, but not from cattle. Culture independent estimates of the number of sulII gene copies obtained by qPCR did not differ between wild life from the two sample sites, while tetW was significantly higher in samples from MPN than from NCA. CONCLUSIONS: Antibiotic resistant bacteria were not more frequently found in ungulates grazing together with cattle than ungulates without this interaction. SIGNIFICANCE AND IMPACT OF THE STUDY: This study did not indicate that transmission of antibiotic resistant bacteria is a frequent event following co-grazing of wild life and cattle.

Prevalence and antibiotic susceptibility of thermophilic Campylobacter isolates from free range domestic duck (Cairina moschata) in Morogoro municipality, Tanzania.

Prevalence and antibiotic susceptibility of thermophilic Campylobacter isolated from free-ranging ducks was determined in Morogoro Municipality, Tanzania. Ninety intestinal contents from ducks were screened for thermophilic Campylobacter using Skirrow's protocol. Of the Campylobacter jejuni isolates, 50 were tested for sensitivity to 12 antibiotics. Overall prevalence of thermophilic Campylobacter was 80%. The prevalence of Campylobacter in adult ducks (91.3%) was significantly (p < 0.05) higher than ducklings (68.2%). The isolation rate of C. jejuni (81.9%) was significantly (P < 0.001) higher than C. coli (18.1%). All C. jejuni isolates were susceptible to streptomycin, nitrofurantoin and amikacin. Forty eight percent, 74% and 82% of isolates were resistant to cefuroxime sodium, tetracycline and ampicillin respectively. Between 20-50% of isolates were resistant to erythromycin, gentamicin, cloxacillin and amoxicillin. Norfloxacin and ciprofloxacin had lower C. jejuni resistance of 10% and 16% respectively. C. jejuni isolates from adult ducks showed significantly higher rates of resistance (p < 0.05) to most antibiotics than did duckling isolates. High prevalence of thermophilic Campylobacter in ducks could be of public health significance in Morogoro municipality. The observed multidrug resistance in this study poses a threat of transfer of antibiotic resistance to human pathogens because of the close contact between ducks and human.

Implementing poultry vaccination and biosecurity at the village level in Tanzania: a social strategy to promote health in free-range poultry populations.

A social strategy was tested for implementing Newcastle disease (ND) vaccination and biosecurity improvements among free-ranging chicken at village level in Tanzania. In addition to training the local poultry vaccinators, data recorders and poultry-keepers, the strategy involved training and empowering leaders at the district, ward and village level. The trainings covered poultry health, management, and marketing of village chickens, with an emphasis on ND vaccination and improving biosecurity against avian influenza (AI), The study sites included villages in one ward in each of three each three districts (Iringa, Mtwara-Mikindani, and Mvomero) of mainland Tanzania. Ninety-six local leaders at district level and 101 leaders at ward levels were trained. In addition, 196 farmers (households) were trained, as well as 86 vaccinators and 26 data recorders. Data recorders were also trained as poultry first aid workers. ND vaccination was conducted by the vaccinators, supervised by their local leaders with technical assistance from veterinarians. A total of 158,343 village chickens were vaccinated in three rounds of vaccination three months apart. The training and empowerment of local leaders and local implementers was the key element for success as it fostered the feeling of local ownership of the program and prevented conflicts with other development activities within the villages. We conclude that most animal health programs will increase their odds of success by involving local leaders and by addressing the current challenges facing the farmers. Further assessment on the usefulness of this approach is needed.

Investigations on the carrier rate of Pasteurella multocida in healthy commercial poultry flocks and flocks affected by fowl cholera.

Twenty flocks of web-footed birds (Pekin and Muscovy ducks and geese) and eight flocks of chickens raised under intensive management were examined for the presence of carriers of Pasteurella multocida. Five hundred and seventy-eight web-footed birds and 240 chickens from healthy flocks, as well as from flocks affected by fowl cholera, were investigated. A total of 135 isolates (80 from healthy flocks and 55 from flocks affected by fowl cholera) were obtained from the pharyngeal and cloacal mucosae after mouse passage (134 isolates) and culture in selective medium (one isolate). Thirty-five percent (7/20) of the flocks of web-footed birds and 38% (3/8) of chicken flocks had birds carrying P. multocida in the pharynx and/or cloaca. Birds from flocks affected by fowl cholera carried P. multocida at a significantly higher prevalence in the mucosa of the cloaca (P < 0.001) compared with the pharynx, while the opposite was observed in birds from healthy flocks. Extended phenotypic characterization confirmed the presence of P. multocida ssp. multocida, P. multocida ssp. septica and P. multocida ssp. gallicida in the flocks examined. P. multocida ssp. gallicida was exclusively isolated from Pekin ducks, while P. multocida ssp. multocida and P. multocida ssp. septica were obtained from chickens as well as web-footed birds. Each flock was shown to be infected by a single phenotypic clone, but some clones were found in more than one flock. A different clone was found in each of four outbreaks of fowl cholera on one of the farms in the preceding 2 years. Two genotypic and phenotypic clones each of P. multocida ssp. multocida and P. multocida ssp. septica were found. This observation indicated that outbreaks are usually clonal and that elimination of P. multocida from infected farms is possible. The results suggest that healthy poultry, in addition to convalescent carriers, may also be carriers of P. multocida. However, the virulence of P. multocida isolates and resistance of carriers to clinical infection needs to be examined. This is the first report of isolation of P. multocida from the cloacal mucosa of apparently healthy domestic poultry. Sampling of the cloaca appeared to be more sensitive for detecting carriers of P. multocida. Although selective medium was used only to a limited extent, the results suggested that mouse inoculation was a more efficient method of isolating P. multocida from poultry than the use of selective media.

Relationships among Pasteurellaceae isolated from free ranging chickens and their animal contacts as determined by quantitative phenotyping, ribotyping and REA-typing.

One hundred and forty-three Pasteurella spp. strains and 10 unclassified strains obtained from free ranging poultry, dogs and cats were investigated by extended phenotypic characterization. One hundred and forty-nine of these strains were selected for further studies using ribotyping and REA-typing to evaluate the role of dogs and cats in Pasteurella multocida transmission. Seven and six type strains were included for comparison in phenotyping and genotyping, respectively. Eleven clusters and six unclustered strains were revealed by phenotyping. Ribotyping outlined 12 clusters and six unclustered strains. A correlation between clusters obtained by phenotyping and ribotyping was demonstrated which indicated that a genetic basis exists for clusters outlined by quantitative evaluation of phenotypic data. Similarities and differences in hosts, phenotype, ribotype, and zone of isolation were demonstrated among Pasteurella strains investigated. Isolates of P. multocida from ducks were shown to be clonal by both phenotyping and ribotyping. These strains were identical to one of the chickens strains. REA-typing, however, showed that the chicken strain was different underlining that exchange of clones of P. multocida between avian species rarely happens under village conditions. Management practise in the villages suggest the potential for exchange of P. multocida between poultry and animals kept in contact. The present findings, however, did not indicate that clones of P. multocida are widely exchanged between poultry and other animal species, even though close contact exists. In the present investigation exchange of clones of P. multocida was only demonstrated among animals belonging to the same species. Caution is drawn to the use of ribotyping as the sole method for epidemiological typing and tracing of P. multocida. The present results also underline the importance of proper phenotyping in the identification of P. multocida and related species.

Occurrence of Pasteurella multocida and related species in village free ranging chickens and their animal contacts in Tanzania.

Investigation was done to determine the presence of Pasteurella multocida and related species in free ranging chickens and ducks, dogs, cats and pigs in three climatic zones (cool, warm and hot) of rural Morogoro, Tanzania. A total of 153 isolates of P. multocida ssp. multocida and related species were obtained by direct culture on blood agar, selective medium and mouse inoculation. P. multocida ssp. multocida was isolated from 0.7% of chickens and 7% of ducks. In dogs and cats, P. multocida ssp. multocida was isolated from 1 and 68%, respectively. One isolate of Pasteurella gallinarum was isolated from a duck. Other species obtained were; P. multocida ssp. septica, Pasteurella stomatis and taxon 16 from dogs and cats, while Pasteurella dagmatis and Pasteurella canis were found in dogs only. Prevalence of P. multocida ssp. multocida was significantly higher (P<0.01) in ducks of the warm zone (22%) than in ducks of other zones (0%). No significant difference was observed between the prevalence of P. multocida ssp. multocida in chickens of the warm zone (2%) and chickens of the cool and hot zones (0%). Extended phenotypic characterization revealed phenotypic similarities between two isolates from chickens and the duck strains. Mouse inoculation appeared to be more sensitive in detecting P. multocida ssp. multocida than blood agar and selective medium. Direct culture on blood agar recovered most of the isolates from dogs. This study has demonstrated for the first time the presence of P. multocida and related species in the village free ranging chickens, ducks, dogs and cats in Tanzania. Other non-classified Pasteurella spp. were also observed in the study, but further characterization is required before the final classification can be made. This paper reports for the first time the isolation of unclassified Pasteurella from dogs and cats in Africa. The results implies that fowl cholera might be occurring in free ranging poultry, and dogs and cats kept in contact might serve as sources of P. multocida to chickens and ducks. Subsequent applications of molecular techniques to analyse the epidemiological relatedness of clones isolated from different host species is indicated.

The effect of concurrent infections with Pasteurella multocida and Ascaridia galli on free range chickens.

Pasteurella multocida and Ascaridia galli are observed with high prevalences in free range chickens in Denmark, but the impact is unknown. A study was carried out to examine the interaction between A. galli and P. multocida in chickens and the impact on production. Five groups, each with 20 18-week-old Lohmann Brown chickens were infected. Group 1 was orally infected with 1000+/-50 embryonated A. galli eggs. Group 2 received 10(4) cfu P. multocida intratracheally. Group 3 was infected with A. galli and subsequently with P. multocida. Group 4 was infected with P. multocida followed by A. galli. Group 5 was the control. The study ran for 11 weeks where clinical manifestations, weight gain and egg production were recorded. Excretion of P. multocida was determined on individual basis and blood smears were made for differential counts. At the end of the study pathological lesions and the number of adult worms, larvae and eggs in the faeces were recorded. The birds were more severely affected when infected with both pathogens compared to single infections with A. galli or P. multocida, respectively. A lower weight gain and egg production was observed with dual infections. A. galli infection followed by a secondary P. multocida infection resulted in more birds with pathological lesions and continued P. multocida excretion. In conclusion a negative interaction between A. galli and P. multocida was observed and it is postulated that free range chickens are at higher risk of being subjected to outbreaks of fowl cholera when they are infected with A. galli.

Helminthosis in local and cross-bred pigs in the Morogoro region of Tanzania.

We investigated the prevalence, burden and types of gastro-intestinal helminths in 424 local and cross-bred pigs kept under different management systems in two climatic zones in the Morogoro region of Tanzania. Coprological examination revealed that 53% of the pigs excreted helminth eggs in their faeces. The median eggs per gram of faeces (EPG) was 500 (range 100-22,000). Local breeds in the Mgeta location with tropical highland climate showed significantly higher prevalence (P < 0.001) and median EPG values (P < 0.001) than the cross-bred animals in the semi-arid area. There was no significant difference in the prevalence (P = 0.90) of helminth infection and egg outputs (P = 0.78) in cross-bred pigs raised under the small-scale and semi-intensive management systems in the semi-arid zone. Piglets showed significantly lower prevalence of helminthosis (P < 0.001) than the weaners, growers and adults in both local and cross-bred animals. Median EPGs of growers and adult animals were significantly higher than those of piglets and weaners (P = 0.006). The prevalences of various helminth species were Oesophagostomum sp. (40%), Ascaris suum (12%), Strongyloides ransomi (9%) and Trichuris suis (5%).

Prevalence of Cryptosporidium spp. oocysts in cattle and wildlife in Morogoro region, Tanzania.

Prevalences of Cryptosporidium spp. oocysts in cattle (n = 486) on five selected farms in Morogoro municipality and three species of herbivorous wildlife (n = 87) from Mikumi National Park, Morogoro, Tanzania, were determined using the modified Ziehl-Neelsen staining technique. Of 486 bovine faecal samples, 5.3% were positive for Cryptosporidium spp. The prevalence of Cryptosporidium was higher in calves less than 3 months of age compared to weaned calves and adults. Cryptosporidium spp. oocysts were detected in both diarrhoeic and non-diarrhoeic animals, but there was a significantly higher prevalence (P < 0.001) of oocyst shedding in diarrhoeic than in non-diarrhoeic animals. Of the 87 faecal specimens from the wildlife species, 36 were from the African buffaloes (Syncerus caffer), 25 from zebra (Equus zebra) and 26 from the wildebeest (Connochaetes gnou). Cryptosporidium spp. oocysts were detected in eight (22%) buffaloes, seven (28%) zebras and seven (27%) wildebeests. Confirmation of the diagnosis was performed using anti-Cryptosporidium monoclonal antibody specific for Cryptosporidium muris. Cryptosporidium parvum and Cryptosporidium baileyi (Pathasure Cryptosporidium test kit).

Eimeriosis in dairy cattle farms in Morogoro municipality of Tanzania.

Coccidial oocysts were detected in 35% of 445 cattle in four medium-scale and 20 small-scale dairy farms in Morogoro municipality, Tanzania. The highest prevalence (56%) was observed in animals aged between 5 and 18 months, whereas lower prevalences were observed in calves (29%) aged between 12 days and 4 months and adults (30%). No coccidial oocysts were detected in calves less than 12 days old. The oocyst output was high in calves, followed by weaners; adults had the lowest oocyst output. The number of oocysts per gram of faeces was significantly higher (P < 0.001) in diarrhoeic animals than in non-diarrhoeic animals, and more so in young calves. Eimeria species infecting the animals included Eimeria bovis (68%) and Eimeria zuernii (57%), Eimeria ellipsoidalis (25%), Eimeria cylindrica (23%), Eimeria auburnensis (22%), Eimeria alabamensis (12%) and Eimeria subspherica (5%). Mixed infections involving two or three species were common. Our findings indicate that eimeriosis is common in cattle in Morogoro municipality.