A qualitative study to explore farmworkers' knowledge, beliefs and preventive practices toward ticks and tick-borne diseases.

INTRODUCTION: Farmworkers are considered a high-risk group for tick-borne diseases (TBDs). This qualitative study aimed to gain an in-depth understanding of the farmworkers' experience, knowledge, health beliefs, information needs, and preventive practices of tick bites and TBDs. METHODOLOGY: A total of nine focus group discussions with 56 farmworkers across eight animal farms in Peninsular Malaysia were conducted between August and October 2013. RESULTS: Farmworkers explained their experience of tick bites, but no one reported TBDs. Many farmworkers indicated that they did not seek any medical treatment. There was a misconception that ticks are solely pathogenic to farm animals. Farmworkers perceived low severity and susceptibility of tick bites, and low self-efficacy of tick bite prevention, however, a group also perceived susceptibility to getting tick bites due to the characteristics of their job. Barriers for prevention were related to the perception and knowledge towards ticks. Farmworkers requested information about TBDs. CONCLUSIONS: This study of farmworkers identified gaps in the knowledge of TBDs, barriers of the tick bite preventive measures and information needs. These findings suggest a need for education programs to improve the knowledge of ticks and TBDs, change health beliefs and address the barriers of tick bite preventive measures.

Ehrlichia and Anaplasma Infections: Serological Evidence and Tick Surveillance in Peninsular Malaysia.

Little information is available on human anaplasmosis and ehrlichiosis in Southeast Asia despite increasing reports of the detection of Anaplasma spp. and Ehrlichia spp. in the ticks. We report herein the serological findings against the tick-borne pathogens in a group of animal farm workers (n = 87) and indigenous people (n = 102) in Peninsular Malaysia. IgG antibodies against Ehrlichia chaffeensis were detected from 29.9% and 34.3% of farm workers and indigenous people, respectively, using commercial indirect immunofluorescence assays. Comparatively, only 6.9% of the indigenous people but none of the animal farm workers were seropositive to Anaplasma phagocytophilum. A polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Anaplasmataceae was used to identify Anaplastamataceae in ticks collected from various locations adjacent to the areas where the serological survey was conducted. In this study, a total of 61.5% of ticks infesting farm animals, 37.5% of ticks infesting peri-domestic animals in rural villages, 27.3% of ticks collected from wildlife animals, and 29.1% of questing ticks collected from forest vegetation were positive for Anaplasmataceae DNA. Sequence analyses of 16S rRNA gene region (238 bp) provide the identification for Anaplasma marginale, Anaplasma bovis, Anaplasma platys, A. phagocytophilum, and Anaplasma spp. closely related to Candidatus Cryptoplasma californiense in ticks. E. chaffeensis DNA was not detected from any ticks, instead, Ehrlichia sp. strain EBm52, Ehrlichia mineirensis and Candidatus Ehrlichia shimanensis are the only Ehrlichia sp. identified from cattle ticks in this study. Further investigation is required to ascertain the occurrence of zoonotic transmission of Ehrlichia and Anaplasma infections in Peninsular Malaysia.

Rickettsial seropositivity in the indigenous community and animal farm workers, and vector surveillance in Peninsular Malaysia.

Rickettsioses are emerging zoonotic diseases that are often neglected in many countries in Southeast Asia. Rickettsial agents are transmitted to humans through exposure to infected arthropods. Limited data are available on the exposure of indigenous community and animal farm workers to the aetiological agents and arthropod vectors of rickettsioses in Peninsular Malaysia. Serological analysis of Rickettsia conorii and Rickettsia felis was performed for 102 individuals from the indigenous community at six rural villages and 87 workers from eight animal farms in Peninsular Malaysia in a cross-sectional study. The indigenous community had significantly higher seropositivity rates for R. conorii (P<0.001) and R. felis (P<0.001), as compared to blood donors from urban (n=61). Similarly, higher seropositivity rates for R. conorii (P=0.046) and R. felis (P<0.001) were noted for animal farm workers, as compared to urban blood donors. On the basis of the sequence analysis of gltA, ompA and ompB, various spotted fever group rickettsiae closely related to R. raoultii, R. heilongjiangensis, R. felis-like organisms, R. tamurae, Rickettsia sp. TCM1, R. felis, Rickettsia sp. LON13 and R. hulinensis were identified from tick/flea samples in animal farms, indigenous villages and urban areas. This study describes rickettsial seropositivity of the Malaysian indigenous community and animal farm workers, and provides molecular evidence regarding the presence of rickettsial agents in ticks/fleas infesting domestic animals in Peninsular Malaysia.

Prevalence and molecular heterogeneity of Bartonella bovis in cattle and Haemaphysalis bispinosa ticks in Peninsular Malaysia.

BACKGROUND: Bartonellosis is an emerging zoonotic infection responsible for a variety of clinical syndromes in humans and animals. Members of the genus Bartonella exhibit high degrees of genetic diversity and ecologic plasticity. The infection is usually transmitted to animals and humans through blood-feeding arthropod vectors such as fleas, lice, ticks and sandflies. This study was conducted to investigate the prevalence of Bartonella species in 184 beef cattle, 40 dairy cattle, 40 sheep and 40 goats in eight animal farms across Peninsular Malaysia. Bartonella-specific PCR assays and sequence analysis of partial fragments of the citrate synthase gene were used for detection and identification of B. bovis. Isolation of B. bovis was attempted from PCR-positive blood samples. Molecular heterogeneity of the isolates was investigated based on sequence analysis of gltA, ITS, rpoB genes, ERIC-PCR, as well as using an established multilocus sequence typing (MLST) method. The carriage rate of B. bovis in ticks was also determined in this study. RESULTS: B. bovis was detected using Bartonella gltA-PCR assays from ten (4.5 %) of 224 cattle blood samples, of which three (1.3 %) were from beef cattle and seven (3.1 %) were from dairy cattle. None of the blood samples from the sheep and goats understudied were positive for B. bovis. Haemaphysalis bispinosa and Rhipicephalus (Boophilus) microplus were the predominant tick species identified in this study. B. bovis was detected from eight of 200 H. bispinosa ticks and none from the R. microplus ticks. Isolation of B. bovis was successful from all PCR-positive cattle blood samples, except one. Strain differentiation of B. bovis isolates was attempted based on sequence analysis of gltA, ITS, rpoB, and ERIC-PCR assay. B. bovis isolates were differentiated into six genotypes using the approach. The genetic heterogeneity of the isolates was confirmed using MLST method. Of the six MLST sequence types identified, five were designated new sequence types (ST23-27), while one (ST18) had been reported previously from Thailand isolates. All except one isolates were segregated into lineage II. A new lineage (IIa) is proposed for a single isolate obtained from a dairy cow. CONCLUSIONS: The current study reported the first detection of B. bovis infection in the cattle and H. bispinosa ticks in Peninsular Malaysia. At least six genotypes of B. bovis were found circulating in the cattle understudied. New MLST sequence types were identified in Malaysian B. bovis isolates. Further study is necessary to explore the zoonotic potential of B. bovis and the vector compatibility of H. bispinosa ticks.

Detection of Schistosoma spindale ova and associated risk factors among Malaysian cattle through coprological survey.

The present study was conducted to determine the occurrence of Schistosoma spindale ova and its associated risk factors in Malaysian cattle through a coprological survey. A total of 266 rectal fecal samples were collected from six farms in Peninsular Malaysia. The overall infection rate of S. spindale was 6% (16 of 266). Schistosoma spindale infection was observed in two farms, with a prevalence of 5.4% and 51.9%, respectively. This trematode was more likely to co-occur with other gastro-intestinal parasites (i.e., Dicrocoelium spp., Paramphistomum spp., strongyle, Eimeria spp. and Entamoeba spp.). Chi-square analysis revealed that female cattle are less likely to get S. spindale infection as compared to male cattle (OR = 0.3; 95% CI = 0.08-1.06; p < 0.05), and cattle weighing lower than 200 kg, were significantly at higher risk than those higher than 200 kg (OR = 5; 95% CI = 1.07-24.79; p < 0.05) to the infection. Multivariate analysis confirmed that among the cattle in Malaysia, the age (cattle with two year old and higher: OR = 21; 95% CI = 2.48-179.44; p < 0.05) and weight (weighing 200 kg and lower: OR = 17; 95% CI = 3.38-87.19; p < 0.05) were risk factors for S. spindale infection among Malaysian cattle.