First study of topical selamectin efficacy for treating cats naturally infected with Brugia malayi and Brugia pahangi under field conditions.

Lymphatic filariae are important human and animal parasites. Infection by these parasites could lead to severe morbidity and has significant socioeconomic impacts. Topical selamectin is a semi-synthetic macrocyclic lactone that is widely used to prevent heartworm infection. Up until now, there were no studies that investigated the efficacy of selamectin in lymphatic filariae. Therefore, we aimed to study the chemotherapeutic and chemoprophylactic efficacies of selamectin use for cats in brugian filariasis-endemic areas in Southern Thailand. To assess chemotherapeutic efficacy of topical selamectin, eight Brugia malayi and six Brugia pahangi microfilaremic cats were treated with a single administration of topical selamectin. For chemoprophylactic efficacy assessment, a single application of topical selamectin was administrated to 9 healthy, uninfected cats. The cats in both groups were subjected to a monthly blood testing for microfilariae and filarial DNA for 1 year. Topical selamectin treatment in B. malayi and B. pahangi microfilaremic cats showed 100% effectivity in eradicating microfilaremia but only 78.5% effectivity in eliminating filarial DNA. In the chemoprophylactic group, selamectin demonstrated 66.7% efficacy in preventing B. malayi infection. Our findings suggest that a single administration of 6 mg/kg topical selamectin given every two months could effectively prevent B. malayi infection. Application of topical selamectin twice a year could block circulating microfilariae. Since there are no treatment guidelines currently available for lymphatic filarial infection in cats, the data obtained from this study could be used to guide the management of brugian lymphatic filarial infection in reservoir cats.

Therapeutic trial of doxycyclin plus ivermectin for the treatment of Brugia malayi naturally infected cats.

Lymphatic filariasis (LF) is one of the neglected tropical diseases which causes permanent and long term disability worldwide. LF is caused by filarial nematode parasites, i.e. Wuchereria bancrofti, Brugia malayi, and B. timori. All available antifilarial drugs currently being used have shown a limited adulticidal activity. Discoveries of endosymbiont rickettsia-like bacterium, Wolbachia in filarial nematodes provided a novel approach for antibiotic use in eradication of filarial diseases. The earlier studies revealed the macrofilaricidal efficacy of doxycycline against filarial nematodes. Chemotherapeutic efficiency of doxycycline has been studied against many filarial parasites, but there are still no therapeutic trials of the drug regimens for B. malayi treatment in naturally infected cats. Thus, this study would be the first attempt to study the efficiency of doxycycline (DOXY) alone or in combination with ivermectin (IVM) for treatment of B. malayi in naturally infected cats. A total of 26 B. malayi-infected cats in the endemic areas were recruited and divided into 3 groups, receiving different treatment regimens; a single dose of ivermectin only (IVM), doxycycline only (DOXY) and a combination of ivermectin and doxycycline (DOXY-IVM). The efficacy of each therapatic regimen was evaluated by detecting the presence of microfilaria using parasitological and molecular techniques monthly up to 2 years after starting the treatment. The IVM treated group had a significant rapid reduction of microfilariae in the first month; however, recurrence of microfilaraemia was observed in some cats. By contrast, the DOXY and DOXY-IVM groups showed a better result with a gradual decrease in microfilariae with no recurrence. These 2 groups were not only virtually deprived of infection but also sustained the sterility of infection through the course of study. These results revealed the advantages of using in B. malayi treatment in cats. Doxycycline showed to have both microfilaricidal and adulticidal effects on lymphatic filariae which maintained the long-term response to control of B. malayi infection in cats.

Rapid Differentiation of Filariae in Unstained and Stained Paraffin-Embedded Sections by a High-Resolution Melting Analysis PCR Assay.

BACKGROUND: Apart from infection with human filariae, zoonotic filariasis also occurs worldwide, and the numbers of cases have been increasing steadily. Diagnosis of intact filariae in tissues or organs depends on histological identification. The morphology of parasites in tissue-embedded sections is poor and shows high levels of homoplasy. Thus, the use of morphological characteristics in taxonomic studies is difficult and may not allow a specific diagnosis. METHODS: Here we report the use of real-time PCR with high-resolution melting analysis (HRM) to detect and identify Brugia malayi, Brugia pahangi, Wuchereria bancrofti, and Dirofilaria immitis in paraffin-embedded sections. Assay specificity was determined using other tissue-dwelling parasites, Angiostrongylus cantonensis, Gnathostoma spinigerum, and Cysticercus cellulosae. We also developed a quick paraffin removal protocol. RESULTS: Both human and animal filariae in formalin-fixed paraffin-embedded sections (FFPES) were diagnosed and identified rapidly, whereas other parasites were negative. There was no difference in the melting temperature of products amplified from filarial DNA obtained from unstained FFPES and Hematoxylin & Eosin-stained sections. Therefore, the DNA extraction protocols developed in this study could be used for real-time PCR with HRM. CONCLUSIONS: We report the successful application of a HRM-PCR assay to differentiate four filarial parasites in FFPES, thus providing the pathologist with an effective alternative diagnostic procedure. Furthermore, the quick paraffin removal protocol developed could shorten the duration and number of steps required for paraffin removal using a standard protocol.

Determination of fecal occult blood in primary schoolchildren infected with Trichuris trichiura.

A correlation of Trichuris trichiura infection and fecal occult blood detection was conducted in 146 primary schoolchildren in Narathiwat Province, Thailand. The Kato-Katz thick smear method was used for determining egg counts and stated as eggs per gram of feces (epg). The number of T. trichiura eggs was categorized as class I (1-499 epg), class 11 (500-4,999 epg), and class III (> 5,000 epg), according to the relation between infection intensity and reduced hemoglobin concentration. Each fecal sample was processed to detect occult blood using a guaiac-based test (Hema-Screen, USA) and an immunochromatographic-based test (HEXAGON OBTI test, Germany). There were 50 schoolchildren without parasitic infection in the control group. Of 96 cases with T. trichiura infection, 85 and 11 children were classified in the class I and class II groups, respectively, but no subjects were in the class III group. Positive occult blood detection results in the control, class I, and class II groups using the guaiac and the immunochemical tests were 0, 3.5, and 9.1% (p=0.19), and 0, 2.4, and 36.4%, (p<0.0001) respectively. This study suggests that T. trichiura infection with an intensity of 500 epg or greater may be associated with intestinal bleeding.

Impact of a filariasis control program on intestinal helminthic infections; a pilot study in Narathiwat Province, Thailand.

This study was conducted in 9 villages located in endemic areas for brugian filariasis in Narathiwat Province, Thailand. Parasitological and anthropometric examinations were cross-sectionally performed to assess the prevalence of intestinal parasitic infections of 539 villagers. Paired stool samples were collected before and after mass treatment for the filariasis control program in 150 participants in order to study the impact of the filariasis control program on intestinal helminthiasis. The results found that 50.3% of the villagers were infected with one or more types of intestinal parasites. Double and triple infections were found in 10.9% and 1.6% of infected individuals respectively. The prevalence of intestinal parasitic infections peaked in the 1-10 year old age-group, which are pre-school and young school-age children. A significant reduction of intestinal helminthic infections in the post-treatment stool sample was observed in the 150 participants who were examined six months after mass treatment. Integrating an intestinal helminthic control program alongside the existing filariasis control program would be an appropriate and cost-effective strategy in the control of intestinal helminths. However, reinfection of parasites was observed.