Knowledge, attitudes and practices regarding rabies and its control among dog owners in Kigali city, Rwanda.

BACKGROUND: Rabies is a zoonotic viral disease that can occur in all warm-blooded animals, including humans. Vaccinating dogs can protect people from contracting rabies. Rabies is a public health threat in Rwanda, but the country does not have information on the epidemiology of rabies. The present study aimed to understand the knowledge, attitudes and practices (KAP) of rabies and its control among dog owners in Kigali city of Rwanda. METHODS: We conducted a cross-sectional survey using a structured questionnaire among 137 dog owners selected from nine administrative study sites. A two-stage random sampling procedure was used to select the participants. Frequency distributions analysis and a series of chi-square tests of associations as well as binary logistic regressions were performed to determine the important factors associated with the response variables. RESULTS: The results showed that 99.5% of respondents knew at least a host susceptible to rabies. Only 22.4% and 21.3% knew that dogs and people can develop rabies, respectively. Nearly 73.6% knew that human rabies can be transmitted through dog-bites and 99% could identify at least a clinical sign of canine rabies. Overall, 81.8% knew that regular vaccination of dogs helps to prevent dog-transmitted human rabies and 43.1% and 26.3% were aware that rabies in humans and in dogs are fatal once clinical symptoms have shown, respectively. Only 69% would observe a dog for 10 days after it bites a man or an animal. Approximately 20.4% were familiar with appropriate cleaning of dog-bites wounds, and 20.6% knew that puppies could receive rabies vaccination before they are three months old. Of those who owned vaccinated dogs, 78% were happy about the cost (US $ ≤ 34) of rabies vaccination. Of all the respondents, 58% had their dogs vaccinated at home by veterinarians while 86% indicated their veterinarians kept rabies vaccines on ice in a cool box. Overall, 53% of the dog owners had sufficient knowledge of rabies, whilst 66% and 17% adopted adequate practices and positive attitudes towards rabies, respectively. Multivariable logistic regression analyses indicated that none of the respondents' sex, educational level, and the length of dog ownership were statistically associated with their knowledge, attitudes and practices of rabies. CONCLUSIONS: This study showed that majority of the dog owners had sufficient knowledge and adopted appropriate practices of rabies. However there exist some knowledge gaps among the dog owners particularly on treatment, transmission and control methods. Therefore, rabies awareness campaign is required to upgrade rabies knowledge of the dog owners on rabies prevention and control in Rwanda.

Infectious pancreatic necrosis virus isolated from farmed rainbow trout and tilapia in Kenya is identical to European isolates.

Infectious pancreatic necrosis virus (IPNV) is an aquabirnavirus that causes serious diseases in a variety of fish species worldwide. It has been isolated from a large number of healthy fresh and marine water fish. Prior to this study, there was no record of the presence of IPNV infection in Kenya. Here, the presence of IPNV in farmed rainbow trout and tilapia was examined in Nyeri County of central Kenya. Head kidney samples taken from five rainbow trout and three tilapia farms and stored in RNALater® were processed by PCR followed by sequencing of a segment A fragment covering nucleotide positions 2,120-2,343 bp. IPNV was detected in all the farms sampled with infection ratios ranging from 0.3 to 0.78 although the infections were not associated with any specific clinical signs of disease. These findings were supported by immunohistochemistry staining of the virus in the kidney and exocrine pancreas of rainbow trout. Sequence alignment and phylogenetic analysis revealed that the Kenyan isolates were identical to European isolates, suggesting a common origin. These findings highlight the need for better biosecurity procedures with more stringent surveillance programmes and control for fish diseases, especially focusing on imported breeding materials to Kenya.

Stinging nettle and neem enhance antibody response to local killed and imported live infectious bursal disease vaccines in indigenous chicken in Kenya.

Immune responses are critical for protection of chickens from infectious bursal disease (IBD). In this study, the antibody response-enhancing effect of drinking water supplementation of 1% stinging nettle and neem on different IBD vaccines and vaccination regimes was evaluated, using 36 (n = 36) specific antibody negative indigenous chicks. The birds were allocated into 3 groups as follows: 1A-C, 2A-C, and 3A-B, while group 3C acted as the unvaccinated non-supplemented control. A local inactivated K1 and imported live attenuated D78 IBD vaccines were given to groups 1A-C and 3A-B at 14 and 28 d of age, respectively. A combination of K1 and D78 vaccines was given 30 d apart to groups 2A and 2B (D78 at 14 and 21 d and K1 at 44 d of age) and on the same d to group 2C at 14 and 28 d of age. Stinging nettle was given in water to groups 1B, 2B, and 2C, and neem to groups 1C, 2A, and 3B. Birds were bled weekly and immune responses monitored using indirect ELISA. Both neem and stinging nettle had antibody response-enhancing effects in groups 1B and 1C, receiving the local inactivated K1 vaccine. There were significant differences (P < 0.05) in antibody titers between groups 1A and 2C. Stinging nettle induced earlier onset of high antibody responses in group 2C and persistent titers (>3.8 log10) from the third week in group 2B. Imported live D78 vaccine induced higher antibody titers compared to the local inactivated K1 vaccine. Groups 2B and 2C receiving a combination of the local K1 and imported live attenuated D78 vaccines had the highest antibody titers. Adoption of stinging nettle supplementation and a prime-boost program involving use of a local virus isolates-derived vaccine is recommended.

Isolation of Infectious Bursal Disease Virus Using Indigenous Chicken Embryos in Kenya.

Infectious bursal disease virus (IBDV) isolates were recovered from outbreaks to initiate activities towards developing a local vaccine strain. Use of indigenous chicken embryos was exploited to determine their potential, promote utilization of local resources for research, and enhance household economic activities. Bursa of Fabricius (BFs) samples from outbreaks shown to be IBDV positive was homogenized and inoculated in 4-week-old specific pathogen-free (SPF) IBDV seronegative white leghorn chicks. The harvested virus was inoculated into 11-day-old indigenous chicken embryos that were IBDV seronegative and passaged serially three times after which they were inoculated into 4-week-old indigenous chicks to test for presence and virulence of propagated virus. Out of 153 BFs collected from outbreaks, 43.8% (67/153) were positive for IBDV antigen and 65.7% (44/67) caused disease in SPF chicks. The embryo mean mortalities were 88% on primary inoculation, 94% in 1st passage, 91% in 2nd passage, and 67% in 3rd passage. After the third passage in embryos all the 44 isolates were virulent in 4-week-old indigenous chicks. The results show that indigenous chicken embryos support growth of IBDV and can be used to propagate the virus as an alternative viral propagating tool for respective vaccine preparation.

Time-course investigation of infection with a low virulent Pasteurella multocida strain in normal and immune-suppressed 12-week-old free-range chickens.

Twelve-week-old indigenous chickens, either immune-suppressed using dexamethasone (IS) or non-immune-suppressed (NIS), were challenged with a low virulent strain, Pasteurella multocida strain NCTC 10322(T), and developed clinical signs and pathological lesions typical of chronic fowl cholera. NIS birds demonstrated much more severe signs of fowl cholera than IS birds. With few exceptions, signs recorded in IS and NIS birds were of the same types, but significantly milder in the IS birds, indicating that immune suppression does not change the course of infection but rather the severity of signs in fowl cholera. P. multocida signals by fluorescent in situ hybridization (FISH) were observed between 1 h and 14 days in the lungs, trachea, air sacs, liver, spleen, bursa of Fabricius and caecal tonsils, while signals from other organs mostly were observed after 24 h. More organs had FISH signals in NIS birds than in IS birds and at higher frequency per organ. Many organs were positive by FISH even 14 days post infection, and it is suggested that these organs may be likely places for long-term carriage of P. multocida following infection. The present study has demonstrated the spread of P. multocida in different tissues in chickens and distribution of lesions associated with chronic fowl cholera, and pointed to a decrease of pathology in IS birds. Since dexamethasone mostly affects heterophils, the study suggests that these cells play a role in the development of lesions associated with chronic fowl cholera in chickens.

[Training program for vitreoretinal surgery in Nairobi, Kenya from 2000 - 2006]. / Trainingsprogramm zur vitreoretinalen Chirurgie in Nairobi, Kenia, in den Jahren 2000 - 2006.

BACKGROUND: After the U. S. Embassy bombing in 1998 in Nairobi, Kenya, a relief operation was immediately initiated by the Department of Ophthalmology of the Ludwig Maximilians University (LMU) Munich, Germany. Surgical devices and material (such as a vitrectomy unit, silicon oil, perfluorodecalin, intraocular lenses, sutures) were supplied to operate on 42 ocular-injured victims of this bombing attack. Apart from these specific operations in Kenya, there is a big need for vitreoretinal surgery in East Africa, as in all other developing countries. Therefore, a vitreoretinal training program was started in 2000 in Nairobi, Kenya. We report about the first 7 years of collaboration between the Ludwig Maximilians University of Munich, Germany and the University of Nairobi in cooperation with the Kenyatta National Hospital, Kenya. MATERIAL AND METHODS: The training program was based on an annual project week in which the author (CLS) carried out vitreoretinal surgery himself and assisted and supervised surgery done by the local Kenyan colleagues at the Kenyatta National Hospital. Within the observation period of the present work (2000 - 2006) the following data were collected: number and kind of surgery, indications, surgeons, grading of eye pathology and the surgical procedure, time to prepare for surgery and duration of surgery. RESULTS: In total, 293 vitreoretinal surgeries were performed during the observation period. Surgeries carried out by the local Kenyan colleagues independently, without intervention of the author, increased from 29.4 % (2000) to 78.6 % (2006). Due to a constant development in the learning process during the project week more severe cases were treated and preparation time for surgery from the nursing side has shortened. CONCLUSIONS: By initiating a project week with high frequency surgery, supported by a highly experienced visiting surgeon, vitreoretinal surgery can be successfully set up in an ophthalmic clinic of a developing country. This approach has some advantages when compared to long-period projects, especially for the purposes of sustainability. However, to be successful it requires systematic planning which is described in this article.

Pasteurella multocida in scavenging family chickens and ducks: carrier status, age susceptibility and transmission between species.

Pasteurella multocida causes fowl cholera, a highly contagious and severe disease in chickens and water fowls. The disease is not well described in less intensive production systems, including scavenging family poultry production in developing countries. P. multocida was isolated from 25.9% of healthy-looking ducks and 6.2% of chickens from free-range family poultry farms and at slaughter slabs at market. On experimental infection with 1.2 to 2.0 x 10(8) organisms of the P. multocida type strain (NCTC 10322(T)), 12-week-old chickens expressed fowl cholera clinical signs significantly more times (372 signs) than those of 4-week-old, 8-week-old and 16-week-old chickens (173, 272 and 187 signs) and more signs were severe. In family ducks the 8-week-old birds expressed clinical signs significantly more times (188 signs) than those of the other age groups (117, 80, and 83 signs, respectively) and severe signs were more frequent. P. multocida transmitted from seeder birds (n=12) to sentinel birds (n=30), which developed clinical signs, and in some cases lesions of fowl cholera allowed bacterial re-isolation, whether infected ducks served as seeders for chickens or chickens served as seeder for ducks. This study has documented the occurrence of P. multocida among healthy-appearing family poultry in a tropical setting, and demonstrated that age susceptibility is highest in 12-week-old family chickens and 8-week-old family ducks when challenged with a low-virulent strain of P. multocida. It has further demonstrated that cross-transmission of fowl cholera may happen between family ducks and chickens, and vice versa.

Monoclonal antibodies identify two neutralization-sensitive epitopes in Besnoitia besnoiti endocytes.

Four monoclonal antibodies were produced against endozoite membrane and cytoplasmic antigens of B. besnoiti. In immunofluorescence antibody tests, three of the clones, designated 2M3C5, 2M1G8 and 2M9G3 recognized antigens restricted to the anterior pole of the endozoites. The fourth clone, 2M9C4, recognized a membrane-associated component in a "beaded" pattern, cytoplasmic granules and extracellular background. The staining characteristics differed from the solid diffuse staining of polyclonal serum. On Western blots of detergent-soluble extracts fractionated under non-reducing conditions in 10% SDS-PAGE gels, mAbs 2M3C5, 2M1G8 and 2M9G3 recognized a common antigen at >200 kDa. Recognition with mAb 2M3C5 was consistently different in intensity and extent. Monoclonal antibody 2M9C4 recognized a single antigen at 75 kDa. The antibodies significantly reduced infectivity of Besnoitia endozoites into cultured cells, demonstrating the potential role of the antigens in the invasion process and raising the possibility of development of a vaccine and diagnostic tests for the disease.

Sensitivity of Listeria species, recovered from indigenous chickens to antibiotics and disinfectants.

BACKGROUND: Resistance of bacteria to antibiotics and disinfectants has been reported widely in the world. Listeria monocytogenes is no exception, although normally it tends to be variably sensitive to many antibiotics and disinfectants. OBJECTIVES: To assess the susceptibility of Listeria isolates recovered from indigenous chickens to commonly used antimicrobials. DESIGN: Nine Listeria isolates recovered from village chickens were tested for sensitivity to commonly used antibiotics and disinfectants and compared with Listeria monocytogenes type strains (L028 and DGH), Staphylococcus aureus NCTC 6571 and Escherichia coli ATCC 25922. SUBJECTS: Nine Listeria isolates. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Susceptibility to eight antibiotics and seven disinfectants. RESULTS: The nine Listeria isolates were sensitive to gentamycin (100%), kanamycin (88.9%), tetracycline (77.8%), cotrimoxazole (66.7%), chloramphenicol (66.7%) and resistant to ampicillin, augmentin and cefuroxime. There was no difference between the antibiotic sensitivity to the various Listeria isolates and Listeria monocytogenes type strains (P > 0.05). The isolates were sensitive to disinfectants; A (100%), B (88.9%), D (77.8%), E (77.8%) but resistant to, CF, and G. There was significant difference between the resistance of Listeria isolates to the various disinfectants at the varied dilutions and the resistance at the recommended user--dilution (P < 0.00293). CONCLUSION: This study has shown that some of the Listeria isolates were resistant to most common antimicrobial agents except gentamycin and disinfectant A. Hence the need to consider this resistance pattern for effective treatment and control of listeriosis.

Carrier status for Listeria monocytogenes and other Listeria species in free range farm and market healthy indigenous chickens and ducks.

BACKGROUND: Listeria organisms are documented to be zoonotic; one of the sources of infection is the domestic fowl where it could occur as in apparent infection. The carriage of Listeria monocytogenes and other Listeria in indigenous birds has not been documented in Kenya. OBJECTIVE: To establish whether healthy looking indigenous chickens and ducks could be carriers of Listeria monocytogenes and other Listeria species. DESIGN: Field survey of indigenous chickens and ducks in three districts of Kenya. SETTING: Embakasi and Dagoreti divisions in Nairobi district; Athi river division in Machakos district; and Ngong division in Kajiado district, in Kenya. SUBJECTS: One hundred and thirty six indigenous chickens and 39 ducks reared under free range scavenging system in Nairobi, Machakos and Kajiado districts, in Kenya, were sampled. METHODS: In surveying the birds, the cloacal and pharyngeal swabs were taken from each bird separately using sterile cotton--tipped applicator swabs. The swabs in saline were transported in a coolbox to the laboratory for bacterial isolation and characterization. INTERVENTIONS: None (only compared farmed and the traded birds). MAIN OUTCOME MEASURES: Isolation of Listeria species and pathogenicity of Listeria isolates. RESULTS: Two Listeria monocytogenes and seven other Listeria species were recovered from the oropharyngeal swab samples of farm and market chickens but none from respective cloacal swabs. No Listeria was recovered from either oropharyngeal or cloacal swabs of farmed duck and slaughter chickens. Traded chickens yielded more Listeria isolates as compared to farmed chickens. CONCLUSION: This study shows that indigenous chickens in Kenya are carriers of Listeria monocytogenes and other Listeria species.

Specific detection of Pasteurella multocida in chickens with fowl cholera and in pig lung tissues using fluorescent rRNA in situ hybridization.

A Pasteurella multocida species-specific oligonucleotide probe, pmhyb449, targeting 16S rRNA was designed and evaluated by whole-cell hybridization against 22 selected reference strains in animal tissues. It differentiated P. multocida from other bacterial species of the families Pasteurellaceae and Enterobacteriaceae and also from divergent species of the order Cytophagales (except biovar 2 strains of Pasteurella avium and Pasteurella canis, which have high 16S rRNA similarity to P. multocida). The potential of the probe for specific identification and differentiation of P. multocida was further detected in formalin-fixed paraffin-embedded lung tissues from experimental fowl cholera in chickens and infections in pigs. In chicken lung tissues P. multocida cells were detected singly, in pairs, as microcolonies, and as massive colonies within air capillaries (septa and lumen), parabronchial septa, and blood vessels (wall and lumen). In pig lung, postmortem-injected P. multocida was detected in the alveoli (lumen and wall), and in both animals the bacterial cells were seen in the bronchi. The results showed that with the oligonucleotide probe pmhyb449, fluorescent in situ hybridization is a suitable and fast method for specific detection of P. multocida in histological formalin-fixed tissues. The test was replicable and reproducible and is recommended as a supplementary test for diagnosis and as a tool in pathogenesis studies of fowl cholera and respiratory tract infections in pigs due to P. multocida.

The comparative antigenic characterization of Newcastle disease virus strains isolated in Kenya and Kazakhstan.

Twenty six Newcastle disease viruses--12 reference strains and 14 strains isolated in Kenya and Kazakhstan--were characterized by means of a large panel of 38 monoclonal antibodies (MAB) directed against all the three envelope proteins: matrix, hemagglutinin-neuraminidase and fusion. The essential distinctions were revealed between the viruses isolated in Kenya and Kazakhstan while the differences amongst the viruses belonging to the same local group were much smaller. The heterogeneity amongst the viruses isolated in Kenya was more expressed as compared to the Kazakhstanian strains.

Comparison of immune responses of two Salmonella gallinarum strains viewed as possible vaccines for fowl typhoid in Kenya.

The immune responses of two S. gallinarum strains, L46 and CN 180, were compared in 15-week-old cockerels. The humoral and cell-mediated immune responses were assayed by means of the indirect haemagglutination test (IHA) and the macrophage migration inhibition test (MIT), respectively. Birds were vaccinated with the two vaccines, respectively, and bled for sera (for IHA) and cells (for MIT) every week up to the seventh week, post vaccination, then every alternate week, three times, and later once every month, for a total period of 37 weeks. Strain L46 was found to induce an immune response that was very similar to that of CN 180. Both gave good humoral and cellular responses.

An epidemic of besnoitiosis in cattle in Kenya.

A total of 17 head of cattle presenting with typical "Elephant skin disease" were isolated from the rest of the herd within a beef ranch for further clinical observation. On physical examination, all the animals had characteristic sclero-conjunctival cysts of Besnoitia besnoiti. In addition, some of the animals had characteristic skin nodules on the legs, ears and back. Histological examination of skin sections revealed typical large Besnoitia cysts. Microscopic examination of crush preparations of skin scrapings revealed crescent-shaped organisms with a more pointed anterior than posterior end (banana-shaped morphology) confirming that the cysts belong to the genus Besnoitia.

Immunosuppression in caprine trypanosomiasis: effects of acute Trypanosoma congolense infection on antibody response to anthrax spore vaccine.

Trypanosoma congolense infected goats were vaccinated with Bacillus anthracis spore vaccine to determine the effect of such infection on the humoral immune response to the vaccine. The anti-anthrax antibody levels were severely depressed in infected goats. When trypanocidal therapy was administered to T. congolense infected goats 14 days after infection they developed antibody levels against Bacillus anthracis similar to uninfected controls.

Haemorrhagic pancarditis in cattle infected with Trypanosoma vivax.

Haemorrhagic pancarditis has been studied microscopically and ultrastructurally. Haemorrhages, oedema, mononuclear cell infiltration, degeneration, fragmentation, atrophy and lysis of myofibres, and extravascular localisation of the parasite were observed.

Market fish hygiene in Kenya.

Vibrio parahaemolyticus was isolated from 53 out of 584 samples (9.1%) of market fish. All strains were Kanagawa negative and were distributed as follows: sea fish 5 out of 370 samples (1.4%), shellfish 48 out of 214 samles (22.4%). Other fish spoilage microflora recovered were: Alcaligenes faecalis, Pseudomonas aeruginosa, Proteus vulgaris, Aeromonas spp. and Vibrio alginolyticus. Total aerobic counts and coliform counts per gram for the lake fish ranged from 2.6 X 10(2) to 6.6 X 10(7) and 10 to 1.0 X 10(2), respectively. Those from marine fish ranged from 1.0 X 10(5) to 8.8 X 10(6) and 2.0 X 10(3) to 1.6 X 10(4), respectively. Counts for marine fish gills alone ranged from 1.4 X 10(5) to 3.4 X 10(8) and 7.2 X 10(2) to 1.4 X 10(7), respectively. No high-temperature (44 degrees) coliforms were recovered from either lake or marine samples.