Prevalence of Methicillin Resistant Staphylococcus aureus(MRSA) in pigs and workers at abattoirs in Trinidad and Tobago

Introduction: Methicillin resistant Staphylococcus aureus (MRSA), a major cause of zoonotic infections, has emerged globally in livestock, particularly pigs. People with occupational contact with food producing animals are at high risk of colonization. The aim of this study was to determine the prevalence of MRSA in pigs and abattoir workers throughout Trinidad and Tobago as well as their resistance to other antimicrobial agents. Methodology: Nasal and skin behind the ear swabs from pigs and nasal swabs from humans were enriched in Mueller Hinton broth with 6.5% sodium chloride, followed by phenol red mannitol broth with 75 mg/L aztreonam and 5 mg/L ceftizoxime. The enriched sample was then plated on both CHROMagar MRSA and Brilliance MRSA. All incubation was at 37ºC for approximately 24 h. Suspect MRSA isolates were confirmed as MRSA using the Penicillin-Binding Protein (PBP2a) test kit and polymerase chain reaction (PCR) to detect the mecA gene. Resistance of the S. aureus and MRSA isolates to 16 antimicrobial agents was determined using the disc diffusion method. Results: Of the 929 pigs and 44 humans sampled, MRSA strains were isolated at a frequency of 0.9% (8/929) and 2.3% (1/44) respectively. All isolates exhibited resistance to one or more of the 16 antimicrobial agents. Conclusions: The study demonstrated that pigs and workers at slaughter houses in Trinidad and Tobago harbour multidrug resistance S. aureus and MRSA. This is of public health significance as occupational exposure of humans can lead to an increased risk of infection and therapeutic failure.

Molecular analysis of Salmonella isolates from poultry by pulsed-field gel electrophoresis

Objective: The study was conducted to determine geneticrelatedness of Salmonella serotypes by pulsed field gel electrophoresis (PFGE). Design and Methodology: A total of 1503 caecal samples of freshly slaughtered poultry were randomly collected from 'pluck shops' across the country. The samples were screened for Salmonella by biochemical, serological and molecular methods. The Salmonella serotypes were analyzed for genetic relatedness for phenotypic antimicrobial resistance, resistance and virulence gene profiles by PFGE generated by digestion with XbaI. Results: Ten different serotypes were detected from all 91 Salmonella isolates. PFGE fingerprinting profiles showed that the Salmonella serotypes in general, were genetically diverse with the detection of a total of 20 PFGE groups. The antibiograms of the isolates were also clearly very variable, which suggest that genotypic antimicrobial resistance may not relate to the phenotypic antibiograms in dendrograms, except for qnrB gene. Results demonstrated a varied spectrum of antimicrobial resistance and PFGE patterns among Salmonella isolates and signify the importance of sustained surveillance of foodborne pathogens in retail poultry pluck shops. Conclusions: The findings provide evidence that poultry from pluck shops are colonized by pathogenic Salmonella harboring antimicrobial resistance genes. The study also reported for the first time in Trinidad, molecular characterization of Salmonella isolates from poultry regarding the relatedness of antibiograms, possession of resistance and virulence genes using their PFGE profiles. The epidemiological surveillance of these serotypes would be necessary to evaluate their possible impact on human health in the country and possibly in the Caribbean region.

Prevention of severe clinical signs and pathological lesions of leptospirosis in experimentally challenged dogs

Objective: To prevent severe clinical and pathological findings of leptospirosis in dogs vaccinated against L. interrogans serovar Copenhageni. Design and Methodology: Two vaccination-challenge experiments involving 22 dogs were performed using a vaccine prepared from formalin-killed cultures of L. interrogans serovar Copenhageni. The dogs were challenged by administering a suspension of 1 x 109 of a virulent strain of serovar Copenhageni (8 mL) at 2 weeks (Study 1: Onset of immunity) and 14 months (Study 2: Duration of immunity) after primary and secondary vaccinations. Each dog was observed for clinical signs of leptospirosis for five weeks post-challenge (PC). Any dog which showed irreversible clinical signs of leptospirosis was humanely euthanized, and a necropsy performed. Results: One (20.0 %) vaccinated puppy in Study 1 showed mild clinical signs (PC) which lasted for one day. Five (100.0 %) non-vaccinated (controls) puppies exhibited irreversible signs of acute severe leptospirosis PC, as well as significant postmortem lesions consistent with leptospiral infection. In Study 2, no clinical signs were exhibited by the vaccinated group of dogs PC, while two (40.0 %) non-vaccinated dogs exhibited mild clinical signs for 2 to 3 days, after which they recovered. Conclusions: The vaccine was successful in protecting vaccinated dogs against acute leptospirosis 2 weeks and 14 months after a vaccination schedule of two doses of the bacterin (primary and booster doses), since all vaccinated dogs were clinically normal after challenge with a virulent inoculum of serovar Copenhageni.

Microbiological quality and potential public health risks of export meat from springbok (Antidorcas marsupialis) in Namibia.

To assess the microbiological quality and safety of export game meat; i) a total of 80 pooled meat samples for aerobic plate count (APC) and Enterobacteriaceae ii) water used in harvesting and processing for microbiological quality and iii) meat and rectal contents for Salmonella spp. and Shiga toxin Escherichia coli (STEC) were evaluated in 2009 and 2010. No differences (p>0.05) in the APCs were observed between the years, but the mean Enterobacteriaceae count for 2009 was 1.33 ± 0.69 log(10)cfu/cm(2) compared to 2.93 ± 1.50 log(10)cfu/cm(2) for 2010. Insignificant Heterotrophic Plate Count (HPC) levels were detected in 9/23 field water samples, while fecal bacterial (coliforms, Clostridium perfringens and enterococci) were absent in all samples. No Salmonella spp. was isolated and all E. coli isolates from meat were negative for STEC virulence genes (stx1, stx2, eae and hlyA), suggesting a negligible role by springbok in the epidemiology of STEC and Salmonella.

Brucellae through the food chain: the role of sheep, goats and springbok (Antidorcus marsupialis) as sources of human infections in Namibia.

A confirmed case of human brucellosis motivated an investigation into the potential source of infection in Namibia. Since domestic animals are principal sources of Brucella infection in humans, 1692 serum samples were screened from sheep, goats and cattle from 4 presumably at-risk farms and 900 springbok (Antidorcas marsupialis) serum samples from 29 mixed farming units for Brucella antibodies by the Rose-Bengal test (RBT) and positive cases confirmed by complement fixation test (CFT). To assess the prevalence of human brucellosis, 137 abattoir employees were tested for Brucella antibodies using the standard tube agglutination test (STAT) and by enzyme linked immunosorbent assay (ELISA). Cattle and sheep from all 4 farms were negative by RBT and CFT but 2 of the 4 farms (Ba and C) had 26/42 and 12/285 seropositive goats, respectively. Post mortem examination of seropositive goats revealed no gross pathological lesions typical of brucellosis except enlarged mesenteric and iliac lymph nodes seen in a single buck. Culture for brucellae from organs of seropositive animals was negative. None of the wildlife sera tested positive by either RBT or CFT. Interviews revealed that besides the case that prompted the investigation, a family and another person from other farms with confirmed brucellosis shared a common history of consumption of unpasteurised goat milk, home-made goat cheese and coffee with raw milk and prior contact with goats, suggesting goats as the likely source of infection. All 137 abattoir employees tested negative by STAT, but 3 were positive by ELISA. The 3 abattoir workers were clinically normal and lacked historical connections with clinical cases. Although goats are often associated with B. melitensis, these studies could not explicitly implicate this species owing to cross-reactivity with B. abortus, which can also infect goats. Nevertheless, these data reinforce the need for a better National Control Programme for brucellosis in Namibia.

Subunit vaccines based on intimin and Efa-1 polypeptides induce humoral immunity in cattle but do not protect against intestinal colonisation by enterohaemorrhagic Escherichia coli O157:H7 or O26:H-.

Enterohaemorrhagic Escherichia coli (EHEC) infections in humans are an important public health concern and are commonly acquired via contact with ruminant faeces. Cattle are a key control point however cross-protective vaccines for the control of EHEC in the bovine reservoir do not yet exist. The EHEC serogroups that are predominantly associated with human infection in Europe and North America are O157 and O26. Intimin and EHEC factor for adherence (Efa-1) play important roles in intestinal colonisation of cattle by EHEC and are thus attractive candidates for the development of subunit vaccines. Immunisation of calves with the cell-binding domain of intimin subtypes beta or gamma via the intramuscular route induced antigen-specific serum IgG1 and, in some cases salivary IgA responses, but did not reduce the magnitude or duration of faecal excretion of EHEC O26:H- (Int(280)-beta) or EHEC O157:H7 (Int(280)-gamma) upon subsequent experimental challenge. Similarly, immunisation of calves via the intramuscular route with the truncated Efa-1 protein (Efa-1') from EHEC O157:H7 or a mixture of the amino-terminal and central thirds of the full-length protein (Efa-1-N and M) did not protect against intestinal colonisation by EHEC O157:H7 (Efa-1') or EHEC O26:H- (Efa-1-N and M) despite the induction of humoral immunity. A portion of the serum IgG1 elicited by the truncated recombinant antigens in calves was confirmed to recognise native protein exposed on the bacterial surface. Calves immunised with a mixture of Int(280)-gamma and Efa-1' or an EHEC O157:H7 bacterin via the intramuscular route then boosted via the intranasal route with the same antigens using cholera toxin B subunit as an adjuvant were also not protected against intestinal colonisation by EHEC O157:H7. These studies highlight the need for further studies to develop and test novel vaccines or treatments for control of this important foodborne pathogen.

Differentiation of Pasteurella multocida isolates from cases of atrophic rhinitis in pigs from Zimbabwe by RAPD and ribotyping.

Atrophic rhinitis in pigs is rarely reported in Southern Africa. To determine the relationship between Pasteurella multocida clones from clinical cases of atrophic rhinitis, twenty-one strains were characterised by selected phenotypic and genotypic methods. Biochemical analysis classified 18 strains as P. multocida subspecies multocida, whilst the remainder were grouped into separate unassigned biotypes. Capsular groups A (16/21) and D (l/21) were found among the isolates by PCR. Four ribotype patterns were obtained following HpaII ribotyping, whilst random amplification of polymorphic DNA (RAPD) revealed three main clusters. However, subclusters were also noted for each RAPD cluster. Our results indicate that RAPD offers a better discrimination of strains than ribotyping and that none of the phenotypic characters were directly related to the genotypic clusters.

Unidentified coryneform bacterial strain from cases of polyarthritis in chickens: phenotype and fatty acid profile.

We report isolation of a strain of fermentative coryneform bacteria from an outbreak of polyarthritis in chickens. This strain could not be assigned to any recognized bacterial taxon because its peculiar phenotype is not yet reported. The strain possessed phenotypic characteristics and fatty acid profile similar to Erysipelothrix but, on the other hand, exhibited temperature-dependent motility like Listeria. We found no evidence of either Mycoplasma synoviae or Chlamydia infection. Details of the phenotype and fatty acid profile of the isolate and measures undertaken to contain the outbreak have been described.

Vaccination to control an outbreak of Mycoplasma crocodyli infection.

Details of a severe outbreak of M. crocodyli infection in farmed crocodiles are reported. The outbreak was suspected to have been precipitated by translocation-related stress on the animals brought from a farm with a known history of M. crocodyli infection. Resorting to the use of an autogenous vaccine proved more effective in alleviating the disease manifestations than antibiotic therapy. Prospects of vaccination in the face of an outbreak are discussed.

Random amplification of polymorphic DNA and phenotypic typing of Zimbabwean isolates of Pasteurella multocida.

Eighty-one isolates presumptively identified as Pasteurella multocida from a variety of diseases in animals in Zimbabwe were subjected to biochemical characterization, capsular typing and RAPD analysis. The majority of isolates (over 80%) were assigned into named taxa and were predominantly P. multocida subsp. multocida and P. multocida subsp. septica, whilst the remainder were unassigned. Serogroup A was predominant among the three capsular types (A, B and D) of P. multocida detected. Three main RAPD clusters and three subclusters were observed among the majority of isolates (93.8%), whilst the remainder was found to be weakly related. Nine different groups of strains with similar RAPD profiles (100% similarity) were also observed. The reference strain of capsular serogroup F clustered with the reference strain of P. multocida subsp. septica, whilst all other serogroups clustered with reference strains of subsp. multocida and gallicida. Notably, serogroups A and D were observed to be closely related to the reference strain of subsp. multocida. The relationship between biotype, capsular type, host origin and disease manifestation was not clear-cut. However, most pig isolates of subsp. multocida clustered together as did most cattle isolates of subsp. multocida. RAPD tended to separate subsp. multocida from septica.

Capsular serogroups of Pasteurella multocida isolated from animals in Zimbabwe.

Pasteurella multocida is isolated from a variety of disease conditions from different animal species in our diagnostic laboratory. In order to determine serogroup distribution among the isolates, an indirect haemagglutination test using glutaraldehyde-fixed sheep red blood cells was employed. A serological examination of 79 isolates revealed that 47/79 were of capsular serogroup A, 11/79 capsular serogroup D, 4/79 capsular serogroup B and 17/79 were untypable strains. None of the isolates belonged to either serogroup E or F. All those from cases of classical pasteurellosis could be grouped, but a significantly high proportion of those which originated from companion animals were untypable. The significance of these results is discussed. This report appears to be the first detailed information on the prevalence of various serogroups of P. multocida in animals in southern Africa.

Pasteurella gallinarum: Zimbabwean experience of a versatile pathogen.

Pasteurella gallinarum-related outbreaks in chickens and African guinea fowls are described. Four outbreaks were recorded in chickens and one in guinea fowls. Periorbital swelling and keratoconjunctivitis were the consistently present clinical signs in all the diseased birds. In several, swollen hocks and wattles were also discerened. Birds which succumbed to the infection showed petechiation in the internal organs and evidence of airsacculitis. Pasteurella gallinarum was isolated from the lesions and also from conjunctival swabs of the apparently healthy in-contact birds. There was no evidence of concurrent infection with Haemophilus, Mycoplasma or Chlamydia. Quinolone therapy when resorted to on one of the farms resolved the clinical signs. Phenotypes of 28 isolates were studied. The results compared well with the Pasteurella gallinarum isolates reported earlier from elsewhere. It was also found that results of xylose fermentation and ONPG test appear to be a variable character. There is no earlier report of P. gallinarum infection in guinea fowls.

Transient infections of goats with a novel spotted fever group rickettsia from Zimbabwe.

A novel spotted fever group rickettsia has recently been isolated from Amblyomma hebraeum ticks in Zimbabwe. In a survey of 172 goat sera collected throughout Zimbabwe the highest prevalence of antibodies reactive with this rickettsia was in the south of the country, the area where A hebraeum is most commonly found. Nine goats were infected using male and female A hebraeum taken from a tick line shown to be infected with the novel rickettsia. By week 3 after infection, seroconversion occurred in all nine goats but no clinical signs of disease could be detected. A leucocytosis due to a mature neutrophilia one to two weeks after infection was the only abnormality. Rickettsaemia was detected only on day 3 after exposure to infected ticks. Immunosuppression failed to induce recrudescence of the rickettsaemia.