Completing the Puzzle: A Cluster of Hunting Dogs with Tick-Borne Illness from a Fishing Community in Tobago, West Indies.

Eight hunting dogs were visited by a state veterinarian on the island of Tobago, Trinidad and Tobago, West Indies, as owners reported anorexia and paralysis in five of their dogs. The veterinarian observed a combination of clinical signs consistent with tick-borne illness, including fever, anorexia, anaemia, lethargy and paralysis. Blood and ticks were collected from each dog and submitted to a diagnostic laboratory for analysis. Microscopic analysis revealed a mixed infection of intracytoplasmic organisms consistent with Babesia spp. (erythrocyte) and Ehrlichia spp. (monocyte), respectively, from one dog, while a complete blood count indicated a regenerative anaemia (n = 1; 12.5%), non-regenerative anaemia (n = 4; 50%), neutrophilia (n = 3; 37.5%), lymphocytosis (n = 2; 25%), thrombocytopaenia (n = 3; 37.5%) and pancytopaenia (n = 1; 12.5%). DNA isolated from the eight blood samples and 20 ticks (16 Rhipicephalus sanguineus and 4 Amblyomma ovale) were subjected to conventional PCR and next-generation sequencing of the 16S rRNA and 18S rRNA gene for Anaplasma/Ehrlichia and Babesia/Theileria/Hepatozoon, respectively. The DNA of Ehrlichia spp., closely related to Ehrlichia canis, was detected in the blood of three dogs (37.5%), Anaplasma spp., closely related to Anaplasma marginale, in two (25%), Babesia vogeli in one dog (12.5%) and seven ticks (35%) and Hepatozoon canis and Anaplasma spp., in one tick (5%), respectively. These findings highlight the need to test both the vector and host for the presence of tick-borne pathogens when undertaking diagnostic investigations. Further studies are also warranted to elucidate the susceptibility of canids to Anaplasma marginale.

Hematological profile of healthy Thoroughbred foals from birth to one month of age in Trinidad, West Indies.

OBJECTIVE: This study aimed to develop a sound database for the hematological reference intervals of thoroughbred foals in Trinidad, West Indies from birth to 1 month of age. ANIMALS: 89 foals. METHODS: Whole blood samples were taken from 89 foals throughout Trinidad at approximately 1 day, 1 week, and 1 month of age. These foals were examined to be classified as healthy or free from disease. Complete blood count (CBC), microscopic analysis of blood smears, and conventional PCR for Theileria equi and Babesia caballi were performed. RESULTS: Of the 89 foals, 67 were deemed healthy and suitable for establishing reference intervals. Foals in this study had lower mean hemoglobin and hematocrit values for all 3 times of sampling when compared to their North American counterparts. Age had a significant effect on hemoglobin, hematocrit, white blood cell (WBC), neutrophil, and platelet counts of the foals from birth to 1 month of age. CLINICAL RELEVANCE: Variations in reference intervals can occur due to differences in demographic, physiological, and environmental factors such as age, gender, breed, and geographical location. Given the changes in the hematological values over time, this study provides clinicians with valuable information that can be used to monitor the health status of newborn foals and detect disease conditions.

Tick-borne pathogens in neotropical animals in Trinidad, West Indies.

BACKGROUND: Ticks are important vectors of many pathogens that have contributed to the morbidity and mortality of humans and domestic animals worldwide. Wildlife species have also been implicated as reservoir hosts of a variety of tick-borne pathogens. The objective of this study was to determine which tick-transmitted pathogens were present in the animals harvested from the forest in Trinidad for human consumption. METHODS: Thin blood smears from 43 neotropical animals were examined microscopically for tick-borne pathogens. Additionally, DNA extraction and PCR amplification of the 16S rRNA gene were used for amplification of Anaplasma and Ehrlichia while the gltA gene was used for Bartonella, and Rickettsia spp. and the 18S rRNA gene for Babesia, Hepatozoon and Theileria species. RESULTS: Pathogen DNA was amplified from four samples (a deer, collared peccary and two agoutis). Sequencing of the amplified products from the deer and collared peccary revealed 99.8% homology to Anaplasma bovis and 98.8% homology to Ehrlichia canis, respectively. Sequences from two agoutis revealed 90.4% homology to Theileria spp. DNA of Hepatozoon spp., Bartonella spp. Babesia spp. and Rickettsia spp. was not detected in any of the screened samples. An incidental finding in this study was the presence of bacteria in the blood of animals. CONCLUSIONS: The results indicate that the DNA of tick-transmitted pathogens is present at a frequency of about 10% in the study population and suggests that neotropical mammals may serve as a source for the potential transmission of tick-borne pathogens to domestic animals and humans. In addition, physicians and hunters should be aware of the symptoms associated with zoonotic tick-borne pathogens so that these infections can be recognised, diagnosed and treated promptly. Bacteria present in carcasses can pose a food safety hazard and hunters should be trained in proper harvesting and handling of carcasses.

A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for Anaplasma/Ehrlichia in Roaming Dogs and Symptomatic Dogs in Trinidad.

This study compared two methods to detect cases of canine ehrlichiosis in a field setting. One method was a polymerase chain reaction for the 16S rRNA gene followed by reverse line blot hybridisation with genera and species-specific probes for Anaplasma/Ehrlichia. The second method was an autologous cell culture of peripheral leucocytes isolated from heparinised blood and maintained in a homologous canine serum in Dulbecco's Modified Eagle medium without antibiotics. The cultures were examined under light microscopy for inclusion bodies after 48 h. Leucocytes were successfully propagated for 20 of the 34 samples submitted for autologous cell culture. Inclusion bodies were observed after cell culture in leucocytes of eight dogs. Two dogs were positive to the Anaplasma/Ehrlichia genera probe and six dogs were positive to the E. canis probe after reverse line blot hybridisation. There was acceptable agreement between reverse line blot hybridisation and cell culture results. Both reverse line blot hybridisation and autologous cell cultures can be used to detect E. canis in subclinical and clinical cases of disease. A definitive diagnosis of E. canis is best achieved by a combination of clinical signs, positive autologous cell culture, and reverse line blot hybridisation results.

A case of Pearsonema eggs in the urine sediment of a cat in Trinidad.

Nematodes of the urinary tract of domestic dogs and cats are a rare occurrence. The discovery of the eggs on urine sediment examination is usually an incidental finding. A twenty-one month old intact queen presented to the Veterinary Teaching Hospital with a history of a serosanguinous vaginal discharge and reddish colour urine for the last ten days. Complete blood count and biochemistry analysis revealed an inflammatory leukogram and a hyperproteinaemia. A urogenital tract infection was diagnosed as haematuria, pyuria, bacteriuria, proteinuria and alkaline urine were evident on urinalysis examination. Microscopic examination of the urine sediment also detected eggs with asymmetrical bipolar plugs characteristic for Pearsonema species. A distended uterus as well as a raised lesion in the mucosal layer of the urinary bladder were observed with ultrasonography. A routine ovariohysterectomy was performed. The cat was also treated with ivermectin and amoxicillin. The cat improved with the eventual resolution of the red colour urine and serosanguinous vaginal discharge.

Survey of ticks of domestic dogs and cattle in three Caribbean islands.

Ticks and the pathogens they transmit can cause high morbidity and mortality in domestic animals. As part of a larger study to determine the tick-borne pathogens infesting domestic animals and wildlife, the aim of this study was to survey the tick species infesting the canine and cattle populations in Trinidad, Tobago and St. Lucia. A total of 1,990 ticks were collected off 179 dogs in Trinidad (n = 163) and Tobago (n = 16) between June 2016 and 2018. Ticks were also collected from cattle throughout Trinidad (n = 1,098), Tobago (n = 306) and St. Lucia (n = 176). Collected ticks were morphologically identified using standard taxonomic keys. Tick-infested dogs were characterized as pets (n = 161) or hunting dogs (n = 18). Only two tick species, Rhipicephalus sanguineus (1,926; 96.8%) and Amblyomma ovale (64; 3.2%), were found on the dogs. A total of 169 (94.4%) dogs and 10 (5.6%) dogs were infested with R. sanguineus and A. ovale, respectively. Three dogs (1.7%) were infested with both tick species. Hunting dogs or those closely associated with them were infested with A. ovale. Rhipicephalus sanguineus was widely distributed throughout both islands, whereas A. ovale was restricted to small foci in three rural settlements in both Trinidad (n = 2) and Tobago (n = 1). Rhipicephalus (Boophilus) microplus (n = 1,404) was the only tick species found in cattle from Trinidad (n = 62) and Tobago (n = 20), whilst R. B. microplus (n = 171) and Amblyomma variegatum (n = 5) were found infesting 14 and two heads of cattle, respectively, in St. Lucia. These preliminary findings will aid in determining whether there are links between ticks and tick-borne pathogens associated with domestic, wildlife species and humans and give further insight into the potential movement of ticks and their pathogens between the human, animal and tropical forest interface.

A comparison of the efficacy of two commercial acaricides (fipronil and amitraz) with Azadirachta indica (neem) on the brown dog tick (Rhipicephalus sanguineus) from canines in Trinidad.

The brown dog tick (Rhipicephalus sanguineus) is prevalent on canids in Trinidad. It is directly (by causing anaemia) and indirectly (by acting as a vector of tick-borne pathogens) responsible for morbidity and mortalities in the canine population. The most commonly used commercial acaricides available to pet owners in Trinidad are amitraz and fipronil. Often, these acaricides may be abused and misused in a desperate attempt to rid pets of ticks. The objective of this study was to compare the efficacy of amitraz and fipronil with the herbal alternative, neem (Azadirachta indica). Triplicate in vitro trials utilizing the Larval Packet Test (LPT) were conducted using three concentrations (low, recommended and high) of fipronil (0.025%, 0.05% and 0.1%), amitraz (0.01%, 0.02% and 1%), neem oil (10%, 20% and 40%) and neem leaf extract (0.25%, 0.5% and 2%) for each trial. Statistical analysis using the mixed-effect Poisson regression analysis indicated that there was a significant difference (p < .05) in the survival of ticks pre-treatment versus post-treatment with amitraz, fipronil and all controls when compared to the neem oil. Fipronil and amitraz caused ≥99% mortality for all concentrations used in this study. Mortalities for neem oil and neem leaf extract ranged from 72.7% to 82% and 38% to 95.3%, respectively, with the greatest percentage of mortalities occurring at the lower concentrations. Neem oil and neem leaf extract can be used as alternative acaricides, and however, they are less efficacious against the brown dog tick than amitraz and fipronil.

Phylogenetic analysis of Theileria equi and Babesia caballi sequences from thoroughbred mares and foals in Trinidad.

The agents of equine piroplasmosis, Theileria equi and Babesia caballi, are endemic in Trinidad, West Indies. While transmission is mainly by ixodid ticks, transplacental transmission of T. equi has also been reported. This disease has contributed to foetal losses as well as morbidity and mortality of neonatal foals and adult horses. Previous 18S rRNA-based phylogenetic studies indicated a noticeable degree of variation within and among B. caballi and T. equi isolates from different geographical regions. The objective of this study was to evaluate the diversity of T. equi and B. caballi obtained from horses in Trinidad by amplifying a region of the 18S rRNA gene. The phylogenetic trees for T. equi sequences obtained from horses in 2006 and 2011-2013 revealed that Trinidad sequences were of genotype A. Additionally, all of the B. caballi sequences from Trinidad were grouped together with other B. caballi sequences of genotype A. However, T. equi sequences from horses in Saint Kitts and Nevis clustered with sequences of genotype C. This study also identified two genotypes of T. equi in the equine population of Brazil. All of the T. equi and B. caballi sequences obtained from horses in Trinidad belong to genotype A and were similar to T. equi and B. caballi sequences of the same genotype that were submitted to GenBank™ databases. Countries in close proximity to Trinidad have T. equi sequences belonging to genotype C; therefore, movement of horses between these countries can introduce a new genotype of T. equi into the equid population of Trinidad.

Novel incidental finding of Hepatozoon canis infection in two dogs of the same household in Trinidad, West Indies.

A five-year old mixed breed bitch was presented to the veterinary clinic in lateral recumbency with a history of anorexia and muscle hyperaesthesia. Examination of the blood smear of this animal revealed the presence of Hepatozoon spp. gamonts in the neutrophils and monocytes with a parasitaemia level of approximately 2%. Complete blood count (CBC) revealed a neutrophilia, and a normocytic normochromic non-regenerative anaemia which were consistent with Hepatozoon spp. infections. Diagnosis was confirmed by polymerase chain reaction (PCR) amplification of the 18S rRNA gene followed by DNA sequencing of the amplicon. Although the other dog in the household appeared asymptomatic, Hepatozoon canis infection was confirmed by both microscopic examination of blood smear and PCR. Both dogs were infested with Rhipicephalus sanguineus ticks. Phylogenetic analysis revealed that the H. canis sequences from these two dogs were similar to those from Venezuela and St Kitts but not Brazil. This is the first reported case of Hepatozoon canis infections in dogs in Trinidad that were confirmed by molecular techniques.

Prospective study investigating transplacental transmission of equine piroplasmosis in thoroughbred foals in Trinidad.

Equine piroplasmosis caused by Theileria equi and Babesia caballi is endemic in Trinidad and Tobago. Transmission occurs by ticks of the family Ixodidae. T. equi can also be transmitted transplacentally; however transplacental transmission of B. caballi is unknown. This study aims to investigate transplacental transmission of equine piroplasmosis from thoroughbred mares naturally infected via the tick vector. Whole blood and serum samples were collected from 117 mares in the fifth month of pregnancy. Blood samples were also collected from each of their foals (89 in total) within the first 36h of birth. Additionally, all foals were observed for clinical signs within 30days post - partum. All samples were examined microscopically for intra-erythrocytic piroplasms. Serum ELISA tests and PCR analysis on whole blood were performed to determine the presence of T. equi and B. caballi. Thirty-four (30.6%) mares and 14 (15.7%) of their foals were seropositive for T. equi. Twenty-seven (24.3%) mares were positive for T. equi by conventional (c) PCR. Real time (q) PCR analysis based on the ema - 1 gene revealed that seven (8%) foals were positive for T. equi. Eighty-nine (76.1%) mares and 38 (42.7%) foals were seropositive for B. caballi. Four (3.4%) mares were positive for B. caballi by cPCR. Three out of the four cPCR positive mares either had resorptions, or stillbirths for that pregnancy. From this study, there is strong evidence that transplacental transmission of B. caballi can occur leading to foetal losses. Six foals (7%) were positive for B. caballi by qPCR. Of these six, four were born to B. caballi seropositive mares. In this study a foal born of a T. equi seropositive mare was 55.7 times more likely to be serologically positive for T. equi than a foal born to a T. equi seronegative mare. Similarly a foal born of a B. caballi seropositive mare was 39.4 times more likely to be serologically positive for B. caballi than a foal born to a mare that was serologically negative for B. caballi at the fifth month of pregnancy. This is as a result of the ingestion of colostrum containing antibodies to these pathogens. Mares should be screened during pregnancy and their foals closely monitored at parturition for evidence of equine piroplasmosis so that treatment can be implemented earlier for a better prognosis.