Alfred Chikweto

Antimicrobial drug resistance as determined by the E-test in Campylobacter jejuni, C. coli, and C. lari isolates from the ceca of broiler and layer chickens in Grenada.

One hundred and twenty five chickens from Grenada, consisting of 77 broilers and 48 layers were examined for carriage of thermophilic campylobacters in their ceca by culture. Seventy nine percent of chickens were positive for campylobacters, with an isolation rate of 93.5% for broilers and 56.3% for layers, the difference being significant. Sixty-four pure cultures comprising 39 Campylobacter coli, 21 Campylobacter jejuni, and 4 Campyilobacter lari isolates were tested for their resistance against 7 antibiotics using the E-test. None of the isolates were resistant to chloramphenicol and gentamicin. Resistance rates to other drugs were: ampicillin, 9.4%; ciprofloxacin, 12.5%; erythromycin, 3.1%; metronidazole, 9.4%, and tetracycline, 50% with MICs of >or=256 microg/mL for tetracycline. There were no significant differences in resistance rates between C. coli and C. jejuni. Multiple resistance to >or=2 drugs was seen in 15.6% of total isolates. All C. lari isolates were resistant to ciprofloxacin, and 3 of 4 isolates had multiple drug resistance. Overall, erythromycin, which is the drug of choice for treatment of Campylobacter infections in humans, is effective in vitro against 97% of chicken isolates in Grenada.

Seroprevalence of Toxoplasma gondii and Concurrent Bartonella Spp., Feline Immunodeficiency Virus, and Feline Leukemia Virus Infections in Cats from Grenada, West Indies

Abstract Toxoplasma gondii and Bartonella spp. are zoonotic pathogens of cats. Feline immunodeficiency virus (FIV) and feline leukemia virus (FeLv) are related to human immunodeficiency virus, and human leukemia virus, respectively; all of these viruses are immunosuppressive. In the present study, the prevalence of antibodies to T. gondi, Bartonella spp., FIV, as well as FeLv antigen were determined in sera from 75 domestic and 101 feral cats (Felis catus) from the Caribbean island of Grenada, West Indies. Using a modified agglutination test, antibodies to T. gondii were found in 23 (30.6%) of the 75 pet cats with titers of 1:25 in 1, 1:50 in 3, 1:400 in 4, 1:500 in 12, 1:800 in 2, and 1:1,600 in 1, and 28 (27.7%) of 101 feral cats with titers of 1:25 in 4, 1:50 in 7, 1:200 in 4, 1:400 in 1, 1:500 in 3, 1:800 in 2, 1:1,600 in 3, and 1:3,200 in 4. Overall, in both pet and feral cats, the seroprevalence increased with age. Antibodies to Bartonella spp. were found in 38 (50.6%) of the 75 pet cats and 52.4% of 101 feral cats. Antibodies to FIV were found in 6 domestic and 22 feral cats. None of the 176 cats was positive for FeLv antigen. There was no correlation among T. gondii, Bartonella spp., and FIV seropositivity.

Existence of Angiostrongylus cantonensis in rats (Rattus norvegicus) in Grenada, West Indies.

The zoonotic rat lung worm, Angiostrongylus cantonensis is endemic to Asia, North America, Africa and Australia. The parasite is expanding geographically and has recently been recorded in some of the Greater Antilles in the northern part of the Caribbean. In this study A. cantonensis is reported for the first time in the Lesser Antilles in one of the southernmost islands, Grenada. Between September 2005 and September 2006, 192 rats (Rattus norvegicus) were trapped throughout the island. The rats were anesthetized, exsanguinated, necropsied and the lungs were fixed whole in 10% buffered formalin, trimmed, processed, cut at 3microm, stained with hematoxylin and eosin and examined microscopically. A total of 45 (23.4%) of the 192 rats examined were found to be infected with A. cantonensis and adult worms were found in the cardiopulmonary system of one of the rats. Microscopically, pulmonic lesions, consisting of pulmonary thrombosis, hypertrophy of pulmonary arteries and granulomatous pneumonia were associated with intralesional adults, larvae and embryonated eggs of A. cantonensis. An incidental finding of variably sized (2-7mm) solitary to multiple cysts containing larvae of Taenia taeniaformis were seen in the livers of 57 rats. This report of A. cantonensis in Grenada provides evidence of the further global expansion of this important zoonotic parasite and the public health implications of this discovery is discussed.

The Phylogeography of Rabies in Grenada, West Indies, and Implications for Control

In Grenada, West Indies, rabies is endemic, and is thought to be maintained in a wildlife host, the small Indian mongoose (Herpestes auropunctatus) with occasional spillover into other hosts. Therefore, the present study was undertaken to improve understanding of rabies epidemiology in Grenada and to inform rabies control policy. Mongooses were trapped island-wide between April 2011 and March 2013 and examined for the presence of Rabies virus (RABV) antigen using the direct fluorescent antibody test (dFAT) and PCR, and for serum neutralizing antibodies (SNA) using the fluorescent antibody virus neutralization test (FAVN). An additional cohort of brain samples from clinical rabies suspects submitted between April 2011 and March 2014 were also investigated for the presence of virus. Two of the 171 (1.7%) live-trapped mongooses were RABV positive by FAT and PCR, and 20 (11.7%) had SNAs. Rabies was diagnosed in 31 of the submitted animals with suspicious clinical signs: 16 mongooses, 12 dogs, 2 cats and 1 goat. Our investigation has revealed that rabies infection spread from the northeast to the southwest of Grenada within the study period. Phylogenetic analysis revealed that the viruses from Grenada formed a monophyletic clade within the cosmopolitan lineage with a common ancestor predicted to have occurred recently (6–23 years ago), and are distinct from those found in Cuba and Puerto Rico, where mongoose rabies is also endemic. These data suggest that it is likely that this specific strain of RABV was imported from European regions rather than the Americas. These data contribute essential information for any potential rabies control program in Grenada and demonstrate the importance of a sound evidence base for planning interventions.

Isolation and RFLP genotyping of Toxoplasma gondii from the domestic dogs (Canis familiaris) from Grenada, West Indies revealed high genetic variability.

Stray dogs are considered as sentinels in the epidemiology of Toxoplasma gondii because they are carnivores and eat a variety of foods, including garbage. In the present study, tissues and sera of 249 stray dogs (Canis familiaris) from Grenada, West Indies were examined for T. gondii infection. Sera were examined for antibodies to T. gondii by the modified agglutination test (MAT); 89 (35.7%) of 249 were seropositive with titers of 25 in seven dogs, 50 in 22 dogs, 100 in 22 dogs, 200 or higher in 38 dogs. Hearts of 76 seropositive dogs were bioassayed in mice. Viable T. gondii was isolated from 12 dogs; these isolates were designated TgDogGr1 to TgDogGr12. These isolates were further propagated in cell culture and DNA isolated from cell culture derived tachyzoites of 12 isolates was genotyped using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The results revealed six genotypes, including ToxoDB PCR-RFLP #1, #2, #3, #7, #13 and #224, with 1, 6, 1, 2, 1 and 1 isolates, respectively. The result supports previous findings that T. gondii population genetics is highly diverse in Grenada.

ISOLATION AND RFLP GENOTYPING OF TOXOPLASMA GONDII FROM THE MONGOOSE (HERPESTES AUROPUNCTATUS) IN GRENADA, WEST INDIES

Little is known of the genetic diversity and epidemiology of Toxoplasma gondii infection in wildlife in Caribbean Islands. The prevalence and genetic diversity of T. gondii in mongooses (Herpestes auropunctatus) was investigated. During 2011 and 2012, 91 mongooses were trapped in different parts of Grenada, bled, euthanized, and examined at necropsy. Antibodies to T. gondii were found in 27 mongooses tested by the modified agglutination test (cut-off titer 25). Muscles (heart, tongue, neck) of 25 of the seropositive mongooses were bioassayed for T. gondii infection in mice. Viable T. gondii was isolated by bioassay in mice from four mongooses with MAT titers of 1:50 in two, 1:200 for one, and 1:400 for one mongoose. The four T. gondii isolates were further propagated in cell culture. Strain typing of T. gondii DNA extracted from cell-cultured tachyzoites using the 10 PCR-restriction fragment length polymorphism (RFLP) markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico revealed one isolate belongs to the Type III (ToxoDB #2) lineage, two to ToxoDB#7 lineage, and one to the ToxoDB #216 lineage. This is the first report of T. gondii isolation and genotyping in H. auropunctatus worldwide.

Spirocercosis in owned and stray dogs in Grenada

The aim of this retrospective study was to estimate the prevalence of Spirocerca lupi and its associated lesions in owned and stray dogs in Grenada. During 2001-2011 necropsies were carried out on 1022 owned and 450 stray dogs at the pathology diagnostic laboratory, School of Veterinary Medicine, St. Georges University, Grenada. Lesions due to S. lupi characterized by focal to multifocal granulomatous esophagitis with aneurysms, mineralized plaques and nodules in the adjacent thoracic aorta were found in 90 (8.8%; 95% confidence interval, 7.1-10.5%) of owned dogs and 64 (14.2%; 95% CI, 11.2-17.6%) of stray dogs. Stray dogs were significantly more affected by spirocercosis than owned dogs (p=0.0022). Of the 90 owned dogs with spirocercosis, 3 dogs had aberrant migration to the thoracic vertebral column with resultant spondylitis; 1 dog each had aberrant migration involving the stomach and the lung. Two dogs had ruptured aorta with hemothorax. Among the 64 stray dogs with spirocercosis, one dog had an esophageal granuloma that transformed into a fibroblastic osteosarcoma; spondylitis due to aberrant migration of S. lupi and hypertrophic osteopathy. We report spirocercosis for the first time in the dogs from a tropical island of Grenada.

Genotypes, Antibiotic Resistance, and ST-8 Genetic Clone in Campylobacter Isolates from Sheep and Goats in Grenada

Rectal swabs from 155 sheep and 252 goats from Grenada were evaluated to determine the prevalence of Campylobacter spp., antibiotic resistance, and multilocus sequence types. Fifteen Campylobacter isolates were obtained (14 C. jejuni and 1 C. coli). The prevalence (3.7%) did not differ significantly between sheep (4.5%) and goats (3.2%). Among the seven antimicrobials tested, resistance was only detected for tetracycline (30.8%) and metronidazole (38.5%). Campylobacter isolates showed no significant difference between sheep and goats for type of antimicrobial resistance or percent of resistant isolates. Twelve of the isolates were successfully genotyped consisting of four recognized clonal complexes and three novel sequence types. Importantly, one isolate from one goat was identified as the C. jejuni sequence type-8, a zoonotic and tetracycline-resistant clone reported to be a highly virulent clone associated with ovine abortion in the USA. Although most samples were from comingled sheep and goat production units, there were no shared sequence types between these two host species. None of the sequence types identified in this study have previously been reported in poultry in Grenada, suggesting sheep- and goat-specific Campylobacter clones in Grenada. This is the first report of genotyping of Campylobacter isolates from sheep and goats in the Eastern Caribbean.

ISOLATION AND RFLP GENOTYPING OF TOXOPLASMA GONDII IN FREE-RANGE CHICKENS (GALLUS DOMESTICUS) IN GRENADA, WEST INDIES REVEALED WIDESPREAD AND DOMINANCE OF CLONAL TYPE III PARASITES.

Abstract The objectives of the present cross-sectional study were to isolate and genotype Toxoplasma gondii in free-range chickens from Grenada, West Indies. Using the modified agglutination test, antibodies to T. gondii were found in 39 (26.9%) of 145 free-range chickens with titers of 25 in 7 chickens, 50 in 6 chickens, 100 in 2 chickens, and 200 or higher in 24 chickens. The hearts of the 39 seropositive chickens were bioassayed in mice; viable T. gondii was isolated from 20 and further propagated in cell culture. Genotyping of T. gondii DNA extracted from cell-cultured tachyzoites using the 10 PCR-restriction fragment length polymorphism (RFLP) markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico revealed 4 genotypes, including ToxoDB PCR-RFLP no. 2 (Type III), no. 7, no. 13, and no. 259 (new). These results indicated that T. gondii population genetics in free-range chickens seems to be moderately diverse with ToxoDB no. 2 (Type III) as the most frequent (15/20 = 75%) compared to other genotypes in Grenada.

Retrospective study of canine heartworm disease with caval syndrome in Grenada, West Indies

Canine heartworm disease caused by Dirofilaria immitis is an important disease of dogs. The aim of this retrospective study was to estimate the prevalence of canine heartworm disease and evaluate cases of caval syndrome in dogs submitted for necropsy in Grenada. Out of 1617 dogs necropsied over a period of 13 years (2001-2013), 249 were found to be infected with D. immitis; giving an overall prevalence of 15.4% (95% confidence interval, 13.6% to 17.1%). There was no significant difference between male and female dogs with respect to canine heartworm infection (p = 0.3). During this period, the annual prevalence of canine heartworm disease was 22% in 2001 before slightly declining to an average of 18% in 2002-2003 and peaking at 26.8% in 2004-2005. From 2006 onwards, annual prevalence rates have steadily been decreasing; reaching the lowest (9%) in 2013. Among the 249 positive cases, 33 (13.2%) of the dogs had caval syndrome. Caval syndrome cases presented with concurrent clinical signs and were associated with cardio-pulmonary and hepatic gross lesions at necropsy. Aberrant migration of D. immitis was also noted in 2 dogs with caval syndrome. This is the first report which presents the findings of canine heartworm disease with caval syndrome in Grenada.