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Clinical and laboratory diagnosis of contagious caprine pleuropneumonia in Qassim region, Saudi Arabia: a comparative study

Elhassan M.A. SAEED ; Salama A. OSMAN

Tropical Biomedicine.2018;35(1):67-75.

Diagnosis of contagious caprine pleuropneumonia (CCPP) in Saudi Arabia mainlydepends on clinical signs and post-mortem findings, in addition to limited usage of latexagglutination test (LAT). In this study, a PCR method specific for detection of Mycoplasmacapricolum subspecies capripneumoniae (Mccp) was used as a direct confirmatory methodand to compare it with clinical signs, necropsy lesions and LAT. During the 2016-2017 year,samples of serum, pleural fluid, lung tissue and nasal swab were collected from 40 goats withclinical signs of CCPP, which were selected from goats brought to the veterinary clinic ofQassim University from 18 goat herds and nine localities. Epidemiological data revealed34.1%, 27.8% and 81.6% morbidity, mortality and case fatality rates, respectively. At necropsy,31 of 40 goats (77.5%) were found with lesions matching those of CCPP. Molecular findingssupported the suitability and applicability of PCR as a reliable method to diagnose andconfirm CCPP directly from clinical samples. The disease was confirmed by PCR in 35 goatsout of 40 (87.5%), 15 herds out of 18 (83.3%) and in all localities. Sera of 32 goats (80%) werefound positive by LAT. Four of the five goats and two of the three herds negative by PCR werealso negative by LAT and necropsy examination. Therefore, PCR sensitivity was considered97.2% (35/36). Compared to the claimed high specificity and sensitivity of the used PCRmethod, diagnosis of CCPP based on clinical signs was found less specific and necropsyexamination and LAT were less sensitive. It was concluded that molecular detection of Mccpdirectly in clinical samples should routinely be used to confirm diagnosis of CCPP in theregion of study, prevent economic impact of wrong diagnosis and to hasten control process.

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Efficacy of Antibiotics (Doxycycline and Kanamycin) against Japanese encephalitis virus infection

Rashmee TOPNO

Tropical Biomedicine.2018;35(1):239-245.

The study evaluated antiviral efficacy of antibiotics- Doxycycline and Kanamycinagainst Japanese encephalitis virus (JEV) infection in vivo. Adult Swiss albino mice (4- 6weeks) were used. Mice were distributed in four groups- control group (A), only drug group(B), JEV infected group (C) and JEV + drug treated group (D). Mice were given intravenousinoculation of JEV strain P20778. Doxycycline was given via intra peritoneal (i. p.) route at 50mg/kg dose. Kanamycin was given to mice via subcutaneous (s. c.) route at 20 mg/kg dose. Alldrug dosages were administered at 24 hr, 48 hr and 96 hr post infection (p.i.) twice a day (BID)for upto 14 days. The mice were monitored for 21 days. The viral load was determined byplaque assay. Viral RNA load and cytokine levels were determined. The infected mice died by8 days of infection. Doxycycline treatment at 50 mg/kg dose after 24 hr p.i. lowered diseaseprogression, prolonging the survival of the animals by a week. Antiviral effect was evidentwith reduction of progeny plaque formation. The plaque formation was reduced at 24 hr p.i.compared to virus group. Doxycycline inhibited viral RNA replication. Doxycycline was ableto moderately modulate proinflammatory cytokines. Kanamycin administration was lesseffective. Thus, the studies demonstrated encouraging results in treatment of JEV infectionby Doxycycline. It indicated that Doxycycline delayed the disease progression. Thus, thefindings suggest that Doxycycline could be used as an adjuvant treatment against JE.

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Parasites of stray and client-owned domestic cats in urban areas in Russia during 2000-2015 years

Tatyana V. MOSKVINA ; Alexander V. TSYBULSKY

Tropical Biomedicine.2018;35(1):267-279.

Cats are commonly infected by many species of parasites that includes helminths,protozoan parasites, mites and fleas. Parasites affect the health of cats and cause morbidityand mortality, especially in young and old animals. Some species such as Ancylostoma spp.,Opisthorchis felineus and Echinococcus spp. are well-known zoonotic parasites worldwide,that high public health risks. Currently of available data on prevalence of feline parasites inRussia, and published studies provide incomplete information regarding the period ofparasitological study, number of cats examined, origin, gender and life condition of examinedanimals. There are no official veterinary guidelines on parasitological examination available.Moreover low quality of veterinary monitoring and inadequate preventive measures has leadto the high rates of environmental contamination by infested cats’ feces containing helminthseggs and protozoan cysts. This paper reviews the knowledge on feline parasites fauna andthe prevalence in Russia. Practical aspects related to diagnosis, treatment, and controls ofparasitic diseases of cats in Russia are discussed.

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Serorevalence of Dirofilaria immitis in dogs and cats in Riyadh City, Saudi Arabia

Omar I. OMAR ; Elgailani A. ELAMIN

Tropical Biomedicine.2018;35(2):531-540.

The aim of present study was to determine the seroprevalence of Dirofilariaimmitis in dogs and cats from Riyadh, Saudi Arabia. Blood samples were collected by cephalicand jugular venipuncture from 294 dogs and 190 cats from Riyadh. Serum samples weretested against circulating D. immitis antigen using DiroCHEK?, SYNBIOTICS Corporation,San Diego, CA 92127, USA and anti-D. immitis antibodies using Green Spring D. immitis IgGantibody ELISA test kit, Shenzhen Lvshiyuan Biotechnology Co., Ltd. Guangdong, China. D.immitis antigens were detected in 7.82% of the dogs and in 1.58% of the cats, while antibodiesagainst D. immitis were detected in 13.61% of the dogs and 2.11% of the cats. In outdoordogs, antigens of D. immitis were detected in 10.33% while antibodies were detected in16.85%. A significant difference in the seroprevalence of D. immitis antibodies in dogs wasreported in the summer compared to the winter (p<0.05). There was no significant differenceobserved in the seroprevalence between males and females dogs. Older dogs (3-6 years)showed higher prevalence of D. immitis antigen compared to younger dogs, in the contraryD. immitis antibody prevalence was higher in younger dogs (1-3 years) compared to olddogs. In cats, the prevalence of D. immitis antigen and antibody was high in males comparedto the females, however, the difference was not statistically significant. There was no effectfor the season and age in the prevalence of D. immitis in cats.

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Hemoprotozoa and Anaplasma spp. in rodents and shrews of Bangladesh

Islam, S. ; Rahman, M.K. ; Ferdous, J. ; Rahman, M. ; Akter, S. ; Faraque, M.O. ; Chowdhury, M.N.U. ; Hossain, M.A. ; Hassan, M.M. ; Islam, A. ; Islam, A.

Tropical Biomedicine.2020;37(No.4):842-851. doi:https://doi.org/10.47665/tb.37.4.842

Hemoprotozoans are important pathogens of animals and humans, among which some species have zoonotic significance. The prevalence of different hemoprotozoa and Anaplasma spp. in larger mammals have been reported from different regions of the world. But, very few studies have been conducted to estimate the prevalence of hemoprotozoa in rodents and shrews of South-East Asia. The study assessed the prevalence of hemoprotozoa and Anaplasma spp. in rodents and shrews of Bangladesh. Blood samples (n=451) were collected from rodents and shrews between June 2011 and June 2013 and July-December 2015 from 4 land gradients of Bangladesh. Giemsa-stained blood smears revealed that 13% of animals were harboring hemoprotozoa (4.7% Babesia spp., 0.67% Plasmodium spp.), and Anaplasma spp. (7.5%). The study may serve as a guide for future hemoparasitic research of rodents and shrews.

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Clone, expression and plasminogen binding property of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis

Li, S. ; Lu, B.P. ; Feng, J. ; Zhou, J.J. ; Xie, Z.Z. ; Liang, C. ; Li, X.R. ; Huang, Y. ; Yu, X.B.

Tropical Biomedicine.2020;37(No.4):852-863. doi:https://doi.org/10.47665/tb.37.4.852

Fructose-1,6-bisphosphate aldolase (FbA), a well characterized glycometabolism enzyme, has been found to participate in other important processes besides the classic catalysis. To understand the important functions of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis (CsFbAs, CsFbA-1/2/3) in host-parasite interplay, the open reading frames of CsFbAs were cloned into pET30a (+) vector and the resulting recombinant plasmids were transformed into Escherichia coli BL21 (DE3) for expression of the proteins. Purified recombinant CsFbAs proteins (rCsFbAs) were approximately 45.0 kDa on 12% SDS-PAGE and could be probed with each rat anti-rCsFbAs sera by western blotting analysis. ELISA and ligand blot overlay indicated that rCsFbAs of 45.0 kDa as well as native CsFbAs of 39.5 kDa from total worm extracts and excretory-secretory products of Clonorchis sinensis (CsESPs) could bind to human plasminogen, and the binding could be efficiently inhibited by lysine analog ε-aminocaproic acid. Our results suggested that as both the components of CsESPs and the plasminogen binding proteins, three CsFbAs might be involved in preventing the formation of the blood clot so that Clonorchis sinensis could acquire enough nutrients from host tissue for their successful survival and colonization in the host. Our work will provide us with new information about the biological function of three CsFbAs and their roles in hostparasite interplay

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Isolation of Beauveria bassiana Pr-11 from Andean orthopterans and its effectiveness against Chagas disease vectors in Peru

Castellanos-Sá ; nchez, P. ; Falconi-Agapito, F. ; Pariona, N. ; Paredes-Esquivel, C.

Tropical Biomedicine.2020;37(No.4):864-870. doi:https://doi.org/10.47665/tb.37.4.864

Chagas disease is endemic to the Americas and is transmitted by blood-feeding kissing bugs. We evaluated the insecticidal potential of a fungus (Beauveria bassiana strain Pr-11) against Triatoma infestans, an important vector in South America. This fungal species was isolated from a locust (Schistocerca piceifrons) that inhabits the Central Andes region of Peru. Ten days post inoculation, this strain induced high insect mortality (97%) at low fungal concentrations (2 × 107 conidia/ml) at 70% relative humidity. The Pr-11 strain outperformed reference strain CCBLE-216 B. bassiana, provided by the Peruvian Ministry of Agriculture. Our results are consistent with previous reports on the virulence of this fungal strain against other insect pests. This is the first study to evaluate an orthopteran-isolated B. bassiana to control Chagas disease vectors. We conclude that strain Beauveria bassiana Pr-11 is effective against Triatoma infestans, resulting in a promising tool to control Chagas disease in Peru and may be used in integrated vector control programs.

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Evaluation of the inhibitory effects of drugs on the growth of Babesia gibsoni using relative quantification real-time PCR

He, W.H. ; Feng, X.X. ; Wu, X. ; Zhai, X.H. ; Li, Y.Y. ; Zhang, B. ; Yao D.W.

Tropical Biomedicine.2020;37(No.4):871-876. doi:https://doi.org/10.47665/tb.37.4.871

To evaluate the inhibitory effects of drugs on the growth of Babesia gibsoni, relative quantification real-time PCR method was developed in this study. The 18S rRNA gene was used as a target gene for the 2–ΔΔCt method analysis. Additionally, chicken RNA was added to the parasitized blood before total RNA extraction. The chicken β-actin gene was selected as an internal control gene for the 2–ΔΔCt method analysis. The 100 µL parasitized blood samples with different percentages of parasitized erythrocytes (PPEs) (3%, 1.5%, 0.75%, 0.375% and 0.1875%) were prepared for relative quantification of B. gibsoni. Regression analysis results revealed significant linear relationships between the relative quantification value and parasitemia. 18S rRNA gene expression was significantly decreased after treatment with diminazene aceturate and artesunate in vitro drug sensitivity test. This result suggested that this relative quantification real-time PCR method can be used to evaluate the effects of drug inhibition.

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The dichotomy of the systemic and local cytokine responses of mucosal leishmaniasis patients in Sudan: A pilot study

Abdelhaleem, A.A. ; Elamin, E.M. ; Elhassan, A.M. ; Nail, A.M. ; Abdullah, N.E. ; Mukhtar, M.M.

Tropical Biomedicine.2020;37(No.4):877-883. doi:https://doi.org/10.47665/tb.37.4.877

Sudanese mucosal leishmaniasis (ML) is a rare clinical form of leishmaniasis and characterized by persistent ulcer of the oral and/or the nasal mucous membranes caused by Leishmania donovani. No data is available about the systemic and local immune responses in mucosal leishmaniasis. This study aimed to measure the systemic and the local cytokines responses of Sudanese ML patients compared to cured cutaneous leishmaniasis patients (Leishmanin skin test positive, LST+ve) and unexposed healthy controls (Leishmanin skin test negative, LST-ve). Six parasitological confirmed ML patients, 7 LST+ve, and 6 LST-ve were enrolled. Systemic Th-1 (IFN-γ and TNF-α), Th-2 (IL-10 and IL-13), Treg (TGF-β1), and inflammatory cytokines IL-6 and IL-8 concentration were measured in the supernatant of whole blood samples following stimulation with live L. donovani promastigotes using ELISA. Local intralesion IL-10, IFN-γ, and IL-13 expression was measured using Real Time PCR. A significant high concentrations of IFN-γ, TNFα, IL-10, TGFβ, IL-6, and IL-8 were detected in the supernatant of stimulated whole blood samples of ML patients compared with the LST+ve and LST-ve controls. Using Real Time-PCR and primers for various cytokines, a significant high expression of TH2 cytokines IL-10 and IL-13 mRNA was detected in contrast to a low TH1 cytokine IFN-γ mRNA in the mucosal lesion. There is a clear dichotomy in the cytokine response during Mucosal leishmaniasis. A significantly high TH1, inflammatory and Treg cytokines response is produced systemically, in contrast to a significant high TH2 cytokines response in the mucosal lesion.

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Bacterial profiling of head lice isolated from the Orang Asli: A first report in Malaysia

Abd Majid, M.A. ; Khoo, J.-J. ; Lim, F.-S. ; Khor, C.-S. ; Loong, S.-K. ; Low, V.-L. ; Lee, H.-Y. ; AbuBakar, S.

Tropical Biomedicine.2020;37(No.4):884-895. doi:https://doi.org/10.47665/tb.37.4.884

This study was carried out to determine from bacterial profiling to the bacterial profiles of head lice among the Orang Asli communities. The head lice were collected from Orang Asli community volunteers. The surface sterilized head lice pools were subjected to genomic DNA extraction while next generation sequencing of the 16S rRNA gene was performed using the Illumina MiSeq platform. Six female and three male head lice identified as Pediculus humanus capitis were collected. A total of 111 368 number of NGS sequencing reads were recorded while another 223 bacterial taxa sequences were obtained. Symbiotic bacteria showed the highest number of reads, with Arsenophonus and Rhodococcus sequences being the most abundant genera in the female and male samples, respectively. The female head lice contained a more distinct microbial diversity. Amongst the pathogenic bacterial species sequences noted were the methicillin-resistant Staphylococcus aureus, Streptobacillus moniliformis, Haemophilus influenzae, Bordetella pertussis and Acinetobacter baumannii. The 16S rRNA genome sequencing revealed a number of rare and pathogenic bacterial species within the head lice of the Orang Asli. The socio-economic practices of the community which involved forest foraging and hunting, and their poor living conditions potentially facilitated the transmission of zoonotic bacterial pathogens, including those found within the head lice. Hence, there is the possibility that the head lice could serve as vectors for the transmission of pathogenic bacteria. This study highlighted the diverse microbial community found within the head lice’s gut of the Orang Asli, with the detection of multiple rare and pathogenic bacteria capable of causing severe infections.

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