Molecular epidemiology of human and animal tuberculosis in Ibadan, Southwestern Nigeria

From 2005 to 2007, Mycobacterium tuberculosis complex (MTC) strains were isolated from cattle, goats and pigs samples collected at the Bodija abattoir and from human samples from tuberculosis patients and livestock traders at the Akinyele cattle market in Ibadan, Southwestern Nigeria. Seventy four isolates obtained from humans (24) and livestock (50) were identified as MTC strains. Thirty two isolates were spoligotyped. Nineteen of these 32 isolates were identified as M. tuberculosis whilst 13 were identified as Mycobacterium bovis. M. bovis was isolated from two humans, whereas M. tuberculosis was isolated from a bovine, a pig and a goat. All the M. bovis isolates identified in this study belonged to the Africa 1 clonal complex. Multiple locus VNTR [variable number of tandem repeats] analysis (MLVA) was carried out on the 74 isolates. Three major clusters were defined. Group A consisted of 24 M. tuberculosis isolates (MLVA genotypes 1-18). One strain was isolated from a bovine and one from a pig. Group B consisted of 49 M. bovis strains (MLVA genotypes 19-48), mainly of cattle origin but also included four goat, nine pig and two human isolates. Group C consisted of a single M. tuberculosis isolate (MLVA genotype 49) obtained from a goat. Spoligotyping and MLVA confirmed it as clustering with the East Africa Indian clade found in humans in Sudan and the Republic of Djibouti. The isolation of three M. tuberculosis strains from livestock raises the question of their epidemiological importance as a source of infection for humans.     

Molecular characterisation of chlamydial isolates from birds

Fifty-one chlamydial isolates from birds collected in Switzerland were classified by amplification and restriction analysis of the 16S-23S rRNA intergenic spacer region as Chlamydophila psittaci. The aim was to characterise a broad panel of chlamydial strains from birds and to apply and verify the methods of classification and differentiation described for chlamydial organisms. Two of the six known avian chlamydial serovars (A and B) were found by serotyping with monoclonal antibodies. One isolate was not typable. Digestion of ompA-PCR amplicons by AluI generated four distinct restriction patterns (genotypes A, B, F and G). Genotypes A and B correlated in most cases to serovars A and B, respectively. One serovar A isolate was verified as genotype B instead of A and one serovar B isolate belonged to genotype A. The non-serotypable isolate was of genotype F and one serovar A generated genotype G. OmpA sequences of one strain of each genotype were determined and compared to data bank entries. Amino acid sequences of genotype A and B strains corresponded well, showing more than 98.0% homology. The homologies of genotypes F and G sequences to genotype A strain were 82.0 and 83.0% respectively.     

Molecular and biological characterisation of Cryptosporidium in pigs

OBJECTIVE: Genetic and biological characterisation of 12 isolates of Cryptosporidium from pigs and comparing them with Cryptosporidium isolates from humans and cattle. DESIGN: Cryptosporidium isolates from pigs were compared with those obtained from human and cattle using rDNA sequence analysis. The infectivity of two of the porcine isolates was determined in neonatal mice and the clinical history of the infected pigs recorded. RESULTS: Pig-derived isolates of Cryptosporidium exhibited two distinct genotypes; a porcine genotype and a bovine genotype, which is common to cattle and other livestock. The porcine genotype did not produce any infection in neonatal mice whereas the bovine genotype did. CONCLUSION: Two distinct genetically and biologically differing strains of Cryptosporidium appeared to be associated with acute diarrhoea in pigs. Whether Cryptosporidium was a primary or secondary pathogen is unclear but warrants further investigation. As the bovine genotype is known to infect humans, the results suggest that pigs can act as reservoirs of cryptosporidial infections for humans and other live-stock. The zoonotic potential of the pig-adapted genotype is uncertain and requires further study.     

Analysis and evaluation of mortality losses of the 2001 African swine fever outbreak,Ibadan, Nigeria

The mortality losses of pigs of various age groups affected by the 2001 African swine fever outbreak in Ibadan Nigeria were analyzed and evaluated. Thirty one thousand nine hundred and sixteen (31,916) pigs on three hundred and six (306) farms reported by the Pig Farmers Association of Nigeria and the State Ministry of Agriculture and Natural Resources were involved. Gross mortality was ninety one percent (91%), while age group mortality ranged from 75.9% (growers), 83.1% (weaners), 91.2% (finishers) and 99.8% (piglets); to 100.0% in gilts, sow and boars. Losses were estimated to worth nine hundred and forty one thousand, four hundred and ninety one dollars, sixty seven cents (US $941,491.67). Highest financial loss was from sows (29.5% of total loss), followed by gilts (16.6%), finishers (15.2%), weaners (10.7%), boars (10.6%), growers (10.6%) and piglets (8.2%). Average mortality loss per farm of $3076.77 was of great financial and socioeconomic consequences for a developing country like Nigeria with a low Gross Domestic Product figures. In conclusion, the need to immediately revisit and take recommended actions on the 1998 Report of the FAO Consultancy Mission to Nigeria on Control and Eradication of an Outbreak of African swine fever in Western Nigeria is stressed.     

Molecular characterization of Cryptosporidium and Giardia isolates from cattle from Portugal

Fecal samples from 291 calves and 176 adult cattle in Northern Portugal were screened for Cryptosporidium and Giardia using a formalin-ethyl acetate concentration method. Acid-fast staining techniques for Cryptosporidium oocyst identification and direct microscopic observation of fecal smears for Giardia cyst identification were performed so as immunofluorescence microscopy examination. Polymerase chain reaction methods were employed to determine the genotype of each isolate. Molecular characterization was performed using amplification and sequencing of the hsp70 and 18SrRNA genes of Cryptosporidium and beta-giardin gene and glutamate dehydrogenase for assemblage determination of Giardia duodenalis. Seventy-four out of 291 calves (25.4%) and 8 out of 176 adult bovines (4.5%) were positive for Cryptosporidium. Forty-one out of 291 calf samples (14.1%) and 1 out of 176 adults samples (0.57%) were positive for Giardia. From the Cryptosporidium positive samples we obtained 63 isolates from calves samples and 7 isolates from adult samples. Additionally, Giardia was isolated in 13 out of 41 positive samples from calves and it was also possible to isolate Giardia from the positive adult sample. Molecular characterization of the Cryptosporidium and Giardia isolates showed us that C. parvum and G. duodenalis assemblage E were the prevalent species. C. parvum may infect humans, representing a potential public health risk. On the other hand, the assemblages B and A2 of Giardia, previously described in humans, were here identified in cattle. Further studies will be needed for determine the importance of cattle as carrier of zoonotic assemblages of G. duodenalis.     

Cryptosporidium hominis Subtypes and Enterocytozoon bieneusi Genotypes in HIV‐Infected Persons in Ibadan,Nigeria

Cryptosporidium and Enterocytozoon are common opportunistic pathogens in HIV+ patients in developing countries, especially those do not have access to antiretroviral therapy. To determine the distribution of genotypes/subtypes of Cryptosporidium and Enterocytozoon bieneusi, faecal specimens were collected from 132 HIV+ persons attending a tertiary hospital in Ibadan, Nigeria. By polymerase chain reaction, eight and ten patients were identified as positive for Cryptosporidium spp. and E. bieneusi, respectively. Seven of the Cryptosporidium specimens were identified as C. hominis, while the remaining one as the new species C. viatorum recently identified in the United Kingdom. DNA sequencing of the 60‐kDa glycoprotein gene showed that the C. hominis belonged to three common subtype families: Ia (in three patients), Ib (in one patient) and Ie (in one patient). In contrast, DNA sequencing of the E. bieneusi internal transcribed spacer products showed the occurrence of genotypes associated with both humans (Peru 8 in one patient, Nig2 in two patients and a new genotype in one patient) and animals (D in one patient and Type IV in five patients). Low CD4+ cell count was identified as a risk factor for both cryptosporidiosis and microsporidiosis.     

Notes on salmonellae isolated from domestic animals in Ibadan,Nigeria

Summary Eight strains of salmonellae isolated from clinical cases in domestic animals in Ibadan, Nigeria are reported. Some of these are reported for the first time either in domestic animals or in the country. The possible danger of resistance to certain antibiotics being developed as a result of such antibiotics being used as food additives, is pointed out.

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Sumario Ocho depas de Salmonellae aisladas de casos clínicos en animales domésticos en Ibadan, Nigeria son reportadas. Algunas de estas son reportadas por primera vez tanto en animales domésticos como en el pais. Se se?ala el posible peligro del desarrollo de resistencia hacia ciertos antibióticos, como resultado de que tales antibióticos estén siendo usados como aditivos de los alimentos.

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Résumé On rapporte ici l’isolement de huit souches deSalmonella à partir de cas cliniques chez des animaux domestiques à Ibadan, Nigéria. Quelques-unes de ces souches sont signalées pour la première fois soit chez les animaux domestiques soit dans le pays. On souligne le danger possible d’une antibio-résistance consécutive à l’emploi d’antibiotiques comme additifs dans les rations alimentaires.

     

Molecular characterisation of Cryptosporidium isolates from pet reptiles

In this study, we investigated the presence of Cryptosporidium in 171 faecal samples from reptiles commonly used as pet animals. These include lizards belonging to the genera Eublepharis, Pogona, Chlamydosaurus, Hemiteconyx, Teratoscincus, Tiliqua, Iguana, and Chamaeleo, snakes of the genera Lampropeltis, Elaphe, Python, Boa and Corallus, and tortoises belonging to the genera Testudo and Kinixys. Cryptosporidium oocysts were detected by immunofluorescence using a commercially available kit and cryptosporidial DNA by amplification of a polymorphic fragment of the 18S rDNA and the HSP70 locus.Cryptosporidium was detected in 38.6% and 25.1% of the samples analysed by immunofluorescence and PCR, respectively. Molecular characterisation of the isolates confirmed that C. serpentis and C. varanii (syn. C. saurophilum) are the main species involved in infection in pet reptiles but also showed the presence of C. parvum and C. muris, as well as other species or genotypes of this parasite including the Cryptosporidium mouse genotype and Cryptosporidium tortoise genotype previously described in reptiles. In addition, a Cryptosporidium sp. was isolated from a chameleon and a python.     

Molecular characterization of Cryptosporidium isolates from pigs in Zaragoza (northeastern Spain)

A total of 142 stool specimens from pigs on 24 farms from the province of Zaragoza (northeastern Spain) were screened for Cryptosporidium spp. Samples were first analysed by routine techniques (formalin-ethyl acetate sedimentation method and modified Ziehl-Neelsen stain) selecting those microscopically positive for genetic characterization. Cryptosporidium species and genotypes were determined by a nested PCR-RFLP technique at the 18S ribosomal DNA locus and sequencing of the PCR-positive secondary products. Cryptosporidium oocysts were microscopically identified in the faeces of 32 pigs (22.5%) from 15 farms (62.5%). Infected animals included 23 weaned piglets (30.7%), 5 fattening pigs (11.9%) and 4 sows (16%). Diarrhoea was not detected in any of the infected pigs. The molecular characterization was successfully performed in 26 samples from 14 farms. Cryptosporidium suis was found in 10 specimens from 7 farms (nine weaned piglets and one sow) and the Cryptosporidium pig genotype II in 16 samples from 10 farms (13 weaned piglets and 3 fattening pigs). Both C. suis and the pig genotype II were concurrently detected on three farms.     

Molecular characterisation of Victorian Newcastle disease virus isolates from 1976 to 1999

OBJECTIVE: To characterise Newcastle disease virus isolates obtained in Victoria from 1976 to 1999 and identify the diversity of FO cleavage signal. DESIGN: RT-PCR using viral RNA extracted from positive NDV allantoic fluid was performed to amplify a segment of the NDV F and HN genes. Molecular characterisation of the nucleotide and amino acid sequences within the FO cleavage site was undertaken. RESULTS: All isolates contained 'avirulent FO cleavage signal sequence of varied amino acid composition. CONCLUSIONS: Molecular characterisation of past and present NDV FO cleavage signal sequences will provide valuable epidemiological information and assist in understanding the genetic origins and relationships of outbreaks.     

Molecular characterization of Cryptosporidium spp. in native breeds of cattle in Kaduna State, Nigeria

Despite numerous molecular epidemiologic studies of cryptosporidiosis in dairy cattle in industrialized countries, there are very few studies on the diversity and public health significance of Cryptosporidium species in native cattle in developing countries. In this study, a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the small-subunit (SSU) rRNA gene was used to detect and identify Cryptosporidium spp. in 194 fecal specimens from 2 to 365 days old calves in 20 White Fulani and Sokoto Gudali herds in Nigeria. Thirty one (16.0%) of the specimens were positive for Cryptosporidium. Restriction digestion of the PCR products showed the presence of Cryptosporidium bovis (7.2%), Cryptosporidium ryanae (4.1%), Cryptosporidium andersoni (2.5%), and concurrent occurrence of C. bovis and C. ryanae (1.5%), and C. bovis and C. andersoni (0.5%). There were no significant differences (p>0.05) in Cryptosporidium infection rates by sex, herd location, management system, breed of calves, or fecal consistency. However, calves 180 days or younger had a higher infection rate of Cryptosporidium than older calves (p=0.034). Likewise, younger calves also had higher occurrence of C. bovis and C. ryanae (p=0.022). The absence of zoonotic Cryptosporidium parvum in the calves studied suggests that native breeds of cattle may not be important in the transmission of human cryptosporidiosis in Kaduna State, Nigeria.     

Identification of Salmonella spp. isolates from chicken abattoirs by multiplex-PCR

The present study was carried out to report the occurrence Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in chicken abattoirs. Samples of feces; feathers; scald, evisceration, and chiller water; and rinse water of non-eviscerated, eviscerated, and chilled carcass were collected from six chicken abattoirs. Salmonella isolates were identified by a multiplex-PCR using three sets of primers targeting the invA, pefA, and sefA gene sequences from Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. Salmonella spp. was detected in 10% (29/288) of the samples, whereas serovars Enteritidis and Typhimurium were identified in 62% (7/288), respectively. The results indicate the need to improve hygiene and sanitary standards in poultry slaughter lines, besides the education of food handlers and information to consumers.     

Castration,other management practices and socio-economic implications for dog keepers in Nsukka area,Enugu state,Nigeria

Structured interviews were used to obtain information from 258 respondents from among 625 people who were selected by stratified random sampling from villages in five of seven local government areas of Nsukka area. Information included gender and literacy status of the respondents, whether the respondents had (between 1990 and 1995) presented any of their dogs for castration, the comparative market value of the dogs, and dog use and owner preference for castrates in performance of such duties. Information on bathing, vaccinations, confinements, use of veterinarians and cultural and religious uses of dogs also were sought. Also, 208 (80%) of the respondents had their dogs castrated.

Of the respondents, 23% were literate, 37% semiliterate and 40% illiterate. Of the 367 non-respondents, 63% were not available during the time of contact and 37% resented dog keeping and therefore refused to talk. About 958 dogs were owned by respondents, of these dogs, 56%, 27% and 17% were females, intact males and castrates, respectively.

There was no association between the respondent’s literacy status and dog-sex preference in performance of such uses as security, hunting, and “economic reasons”. The three most-important reasons for dog keeping were security, pet and hunting. However, the use of castrates for security was favoured by most keepers irrespective of literacy status. Most of the respondents agreed that dogs are used as gifts and 72% of the respondents agree that dog meat is a protein source. Castration should be encouraged because of its market value. Usage was low of veterinary services, confinement, and bathing of dogs.     

Studies on the blood parasites of pigs in Ibadan,Nigeria

The blood parasites in the blood of local pigs kept under free range, and exotic pigs kept under intensive management in Ibadan were investigated. The species found were Babesia trautmanni, B. perroncitoi, Eperythrozoon suis and E. parvum. Eperythrozoon suis was the most predominant in both breeds. Most of the local pigs carried mixed infections, while the same number of the exotic pigs carried single and mixed infections. Mixed infection of individual pigs with all four blood parasites was common. The parasitaemia was generally low in pigs of both breeds.     

Molecular detection of Cryptosporidium spp. infections in water buffaloes from northeast Thailand

The objectives of this study were to determine the individual and herd-level prevalence and genotype of Cryptosporidium and to identify putative risk factors associated with Cryptosporidium spp. infections in water buffaloes in northeast Thailand. Fecal samples from 600 water buffaloes of 287 farms in six provinces were collected and tested using DMSO-modified acid-fast staining and polymerase chain reaction. The overall prevalence of Cryptosporidium infections in buffaloes was 5.7 and 8.7 % among individual animals and herds, respectively. The provinces with highest infected Cryptosporidium were located in the Sakon Nakhon Basin in the northern part of the region. In addition, higher herd prevalence was observed among farms with more than five buffaloes (30 %) than those with five or less animals (16.2 %). Thirty (88.2 %) of the 34 Cryptosporidium-positive samples were Cryptosporidium parvum and four (11.8 %) were Cryptosporidium ryanae.     

Molecular characterisation of avian Pasteurella multocida isolates from Australia and Vietnam by REP-PCR and PFGE

A total of 95 isolates of Pasteurella multocida were analysed by pulsed field gel electrophoresis (PFGE) using the enzyme ApaI, including 73 avian isolates from Australia and 22 from Vietnam. The majority of field isolates were capsular Type A, with the predominant somatic serovars of 1, 3, 4 and 3,4. Twenty-one distinct profiles were evident among the Australian isolates, with only 3 profiles observed among the 22 P. multocida strains isolated from Vietnam. Within the Australian isolates, related and unrelated outbreaks could be identified by PFGE. These results correlated well with previously published studies, with greater discrimination shown by PFGE. Repetitive extragenic palindromic sequence PCR (REP-PCR) analysis of representative isolates from PFGE classifications yielded 21 profiles, with most of the subgroups in accordance with PFGE analysis. While REP-PCR was shown to be less discriminating than PFGE, the epidemiological relatedness of strains compared favourably between the techniques. Thus, the ease and rapidity of REP-PCR while maintaining a high level of differentiation, supports the use of REP-PCR as a competent alternative to the more labour-intensive PFGE system for strain identification and epidemiological studies of avian P. multocida.     

Occurrence of methicillin-resistant staphylococci in the pig-production chain in Ibadan,Nigeria

Staphylococcus species colonises humans and animals and is a major food contaminant with public health significance. Here, we assessed the occurrence of methicillin-resistant staphylococci (MRS) in the pig-production chain in Ibadan, Nigeria. Nares of 120 pigs and 10 farmers were sampled with sterile swabs whilst 54 pork samples were collected from a retail slaughterhouse. Staphylococcus species were isolated using enrichment, cefoxitin–aztreonam selective broth and Mannitol salt agar. Isolates were tested for susceptibility to cefoxitin (30 μg), oxacillin (1 μg) and vancomycin (30 μg). Methicillin-resistant staphylococci isolates were characterised using conventional biochemical tests. From 184 samples, 364 staphylococcal isolates were obtained. Amongst the 54 pork samples, 44.0% were contaminated with Staphylococcus species. Overall, 9 (2.5%) MRS were obtained and presumptively identified as Staphylococcus xylosus (n = 3), Staphylococcus sciuri (n = 3), Staphylococcus warneri (n = 2) and Staphylococcus cohnii (n = 1). There was no relationship between the prevalence of MRS between pigs and pig handlers in the farms, but Farm 2 had the highest frequency of 66.7% (p < 0.05). Piglets had the highest prevalence of 66.7% (p < 0.05) whilst MRS was absent in workers and pork samples. This study raises concerns about the cross-contamination of staphylococci in the food chain. Constant surveillance is imperative to ensure food safety.