Veterinary Parasitology

Ivermectin is considered to be a wonder drug due to its broad-spectrum antiparasitic activity against both ectoparasites and endoparasites (under class of endectocide) and has multiple applications in both veterinary and human medicine. In particular, ivermectin is commonly used in the treatment of different kinds of infections and infestations. By altering the vehicles used in the formulations, the pharmacokinetic properties of different ivermectin preparations can be altered. Since its development, various vehicles have been evaluated to assess the efficacy, safety, and therapeutic systemic concentrations of ivermectin in different species. A subcutaneous route of administration is preferred over a topical or an oral route for ivermectin due to superior bioavailability. Different formulations of ivermectin have been developed over the years, such as stabilized aqueous formulations, osmotic pumps, controlled release capsules, silicone carriers, zein microspheres, biodegradable microparticulate drug delivery systems, lipid nanocapsules, solid lipid nanoparticles, sustained-release ivermectin varnish, sustained-release ivermectin-loaded solid dispersion suspension, and biodegradable subcutaneous implants. However, several reports of ivermectin resistance have been identified in different parts of the world over the past few years. Continuous use of suboptimal formulations or sub-therapeutic plasma concentrations may predispose an individual to resistance toward ivermectin. The current research trend is focused toward the need for developing ivermectin formulations that are stable, effective, and safe and that reduce the number of doses required for complete clinical cure in different parasitic diseases. Therefore, single-dose long-acting preparations of ivermectin that provide effective therapeutic drug concentrations need to be developed and commercialized, which may revolutionize drug therapy and prophylaxis against various parasitic diseases in the near future. The present review highlights the current advances in pharmacokinetic modulation of ivermectin formulations and their potent therapeutic applications, issues related to emergence of ivermectin resistance, and future trends of ivermectin usage.

Sequence analysis of the cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei isolated from goats and rabbits in East Java, Indonesia, Veterinary World, 12(7): 959-964. Abstract Aim: This study aimed to sequence the Cytochrome c oxidase (COX-1) gene sequence from mitochondrial DNA of Sarcoptes scabiei isolated from Lamongan goats and Mojokerto rabbits, align it with DNA isolated from Zi'gong rabbit (GenBank accession No. EU256389.1), and produce a phylogenetic analysis of S. scabiei COX-1 gene. Materials and Methods: S. scabiei mites were obtained from goats and rabbits, and DNA was extracted using QIAamp DNA Mini Kit. The forward and reverse primer sequences were designed based on the DNA sequence of an S. scabiei COX-1 gene isolated from the Zi'gong rabbit (5'-TCTTAGGGGCTGGATTTAGTATG-3' and 5'-AGTTCCTCTACCAGTTCCAC-3', respectively). To confirm sequencing output, the sequence resulting from the reverse primer was inverted and aligned to the sequence from the forward primer using Clone Manager Professional Version 9 for Windows (Scientific & Educational Software; http://www.scied.com). This alignment was subsequently used to build a phylogenetic tree, using the Neighbor-Joining method, in the MEGA6 program (https://www.megasoftware.net/). Results: Polymerase chain reaction (PCR) products from S. scabiei isolates from Lamongan goats and Mojokerto rabbits produced bands of around 290 bp with 2% agarose gel electrophoresis. Comparing the DNA sequences of the S. scabiei COX-1 gene with those isolated from Lamongan goats and Mojokerto rabbits showed 99% homology. Conclusion: PCR products of the S. scabiei COX-1 gene isolated from Lamongan goats and Mojokerto rabbits were around 290 bp long. The sequences had more than 99% homology. The sequences of the COX-1 gene of S. scabiei from Lamongan goats and Mojokerto rabbits were relatively close to the sequence of the gene in S. scabiei obtained from various hosts according to National Center for Biotechnology Information data.

Aim: The current study was designed to isolate and characterize Toxocara vitulorum glycoprotein antigens and then to evaluate its potency in accurate diagnosis of toxocariasis.

Materials and Methods: T. vitulorum glycoprotein fractions were isolated using Con-A affinity chromatography. The fractions characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblot assay. Mass spectrometric analysis was used for identification of proposed structure of the N-acetylglucosamine (GlcNAc) fraction. Enzyme-linked immunosorbent assay (ELISA) was used to assess the diagnostic potential of the isolated fractions.

Results: Surface of T. vitulorum adult worm revealed two glycoprotein fractions rich in glucose (Glc) and GlcNAc. Three bands of molecular weight 212kDa, 107 kDa, and 93 kDa were detected in Glc fraction by SDS-PAGE. These bands were also detected in GlcNAc fraction with an additional band of 49 kDa. GlcNAc fraction showed more diagnostic potency of calves' toxocariasis; 79% than Glc fraction; 46.9% by indirect ELISA. The additional band of 49 kDa in GlcNAc fraction is probably responsible for its higher diagnostic potentials. Western blotting verified the immunoreactivity of the Glc and GlcNAc isolated fraction as they reacted with calves sera infected with toxocariasis. The proposed structure of GlcNAc fraction was Ser-Meth-Arg-O-methylated GlcNAc.

Conclusion: GlcNAc-rich fraction of T. vitulorum can be successfully utilized in the diagnosis of calves' toxocariasis.

Free pre-tetrathyridia of Mesocestoides sp. are described, for the first time, from samples obtained from the coelomic cavity of a ground skink, Scincella lateralis, from Oklahoma. Closer examination of these early-stage tapeworms revealed they were transitional metamorphosis stages between a post-hexacanth procercoid form to the full metacestode of Mesocestoides. A series of transitional stages was found that span the full period of sucker and apical organ development. However, we did not see any fully developed tetrathyridia, i.e., having classic Mesocestoides morphology but with the apical sucker absent following developmental atrophy. This is the first time that metamorphic pre-tetrathyridial stages of a Mesocestoides sp. have been reported in vivo from a natural infection. These observations corroborate earlier reports of such stages of Mesocestoides vogae developed in vitro, though the previously reported isolate of M. vogae is asexually proliferative, and the species from the present study showed no sign of asexual proliferation. The fact that these immediately post-hexacanth stages can occur in a single lizard intermediate host may suggest that Mesocestoides spp. might develop through a simple 2-host life cycle rather than an obligate 3-host cycle that has been speculated to occur by most previous authors. DNA sequence comparisons and phylogenetic analyses based on mitochondrial 12S, cox1, and nad1 genes have demonstrated that our specimens from S. lateralis represent a species clearly distinct from all previously sequenced Mesocestoides and closely related to 2 forms from domestic dogs and Channel Island fox in California previously published as Mesocestoides sp. C.

Ultrastructure of eggs of the cestode Gyro-cotyle urna Grube and Wagener 1852 with fully formed shells and early embryonic stages is described for the first time. Multiple techniques used included cryo-methods of high-pressure freezing with freeze substitution combined with field emission scanning electron microscopy and transmission electron microscopy (TEM). After 20 days of in vitro maturation, eggs are composed, from the exterior to the interior, of a thick egg shell of vitelline origin, an early embryonic envelope, a ring of numerous large vitellocytes, and a group of differentiating blastomeres situated in the center, forming the early presumptive larva. The thick resistant egg shell is similar to that reported from other polylecithal neodermatan flatworms. The embryonic envelope is poorly formed at this stage and not yet divided into inner and outer envelopes as occurs in all neoder-matans described to date. Vitellocytes are separate, not forming a syncytium, and are filled with diverse vitelline materials as well as lipid droplets and associated endomembrane components having ultrastructure consistent with macroautophagy systems seen among many animals in apoptotic or autophagic cells. These results largely corroborate the basic observations and interpretations of previous TEM studies on other neodermatans and provide insight into a powerful suite of cryo-techniques that can enhance our work with these challenging subjects.

Goat farming is the great economic importance as a major source of income for small and the landless farmers in rural areas. A study on prevalence of helminths of goat in relation to age, sex, seasonal, tehsil and month-wise was studied at Udaipur district, Rajasthan

Aim: The objective of this research was to determine the species and strains of Leucocytozoon caulleryi and study the phylogenetics of L. caulleryi of broiler chickens in endemic areas in Indonesia. Materials and Methods: Blood samples were collected from broiler chickens originated from endemic area in Indonesia, i.e., Pasuruan, Lamongan, Blitar, Lumajang, Boyolali, Purwokerto, and Banjarmasin in 2017. Collected blood was used for microscopic examination, sequencing using BLAST method to identify the nucleotide structure of cytochrome b (cyt b) gene that determines the species, and the phylogenetics analysis of L. caulleryi that infected broiler chickens in endemic areas in Indonesia, using Mega 5 software. Results: The results showed that Plasmodium sp. and L. caulleryi were infected broiler chickens in endemic areas in Indonesia. L. caulleryi in one area had very close phylogenetic relations with those in other areas. The genetic distance between L. caulleryi taxa from various endemic areas is very close (<5%). Conclusion: There is a very close phylogenetics among strains of L. caulleryi that infected broiler chickens in various endemic areas in Indonesia.

A three months Toxologic pathology study of piperazine hydrate in male wild pigeons by oral intubation were done. The study was done at three dosage levels as low (0.25 ml), intermediate (0.50 ml) and high (1 ml) with untreated control as fourth group. Clinical observation of treated pigeons did not show significant changes only the birds appeared to be quite after dosing for short time. Macroscopically no obvious changes could be detected. Microscopically, minimal diffuse vacculation of hepatocytes and varying number of parenchymal foci of inflammatory cells, mostly mononuclear cells. In treated birds a renal lesions were characterized by dilated cortical tubules mostly the proximal convoluted tubules with different levels. Heart showed foci/areas of infiltrating mononuclear cells between myocardial muscle cells with occasional vacuolated myocardial muscles cells. The pancreas showed a histopathological changes which were restricted to endocrine islets of langerhanse, which showed evidence of degeneration characterized by vacculation. The nervous system, there was no histopathological changes could be seen in cerebrum, but, vacculation of perking cells was noticed in cerebellum in some of treated birds, while, the most significant histopathological changes were in spinal cord as varying numbers of degenerate/vacuolated nerve fibers at all dosage levels but most severe at high dose levels, the other changes but to less extents were in sciatic nerve as there were only few to occasional degenerate/vacuolated nerve fibers with less severity as they were on the way of recovery.

Heamonchus contortus (order strongylida) is a common parasitic nematode infecting small ruminants and causing significant economic losses worldwide. H. contortus are consider the most economically important parasite which infect cattle, goats and sheep occur in nearly all subtropical and temperate areas of the world. This study was conducted to detected the histopathological changes in the abomasum of the infected sheep in Basrah city and found different changes, like, a stomach glandular region with fibrosis at the base of lamina properia, Infiltration of mononuclear cells at the base of lamina properia, a debris of parasite with completely fibrosis in glandular tissue of abomasum. Some examined infected abomasum showed congested blood vessels in the sub mucosa and, Degeneration of mucus gland associated with infiltration of mononuclear cells, Edema in the lamina properia. It can be seen the part of parasite penetrated mucosa and submuocosa surrounding with inflammatory cells, while, focusing on part of parasite in submuocosa with inflammatory cells.

A study was conducted in sheep on Canadian farms to describe the relationship between packed cell volume (PCV) or fecal egg counts (FEC) and subjective clinical parameters that may indicate the severity of parasitic gastroenteritis. Twenty-one farms in Ontario (ON) and 8 farms in Quebec (QC) were purposively selected and visited during April-May (spring) and August (summer) 2007. At each farm visit, blood and fecal samples were collected from 10 ewes and 10 female lambs; body condition score (BCS), dag score (DS), fecal consistency score (FCS) and FAMACHA score were recorded for all sampled sheep. Packed cell volume was determined for all blood samples, and FEC were performed for all fecal samples. Summary statistics and simple correlations were performed for the parameters recorded. Two mixed models with random effects at the farm level were developed; one using PCV as the response variable and another using the natural log of eggs per gram of feces (lnEPG). Finally, the residuals ...

Trypanosoma evansi, a haemoflagellate, causes &quot;surra&quot; an important chronic wasting disease of a wide range of wild and domestic herbivorous and carnivorous animals including cattle, buffaloes, camels, horses, etc. The untreated recovered animal can act as a carrier without exhibiting the disease symptoms and can be a source of infection to healthy animals. The diagnosis and subsequent treatment of the carrier animals is helpful to curb the disease. As the parasitaemia in carrier animals is very scanty, the conventional blood smear examination, which is widely practiced in the field, cannot detect such condition. For this purpose improved diagnostics are very much useful for mass sero-screening test such as ELISA. In the present study, the VSG of T. evansi was expressed in prokaryotic system (E. coli) and thereafter its immunoreactivity has been evaluated in immuno blot and enzyme immuno assay. The expressed protein showed 95.6% sensitivity, 98.0% specificity and 0.93 Cohen...

Coccidiosis merupakan penyakit protozoa di saluran pencernaan yang disebabkan oleh genus Eimeria. Penyakit ini dapat menyerang pada golongan unggas, mamalia maupun ruminansia. Infeksi klinis maupun subklinis menyebabkan gangguan pada konversi pakan sebab protozoa tersebut akan melakukan tindakan invasif pada epitel usus. Perubahan konversi pakan memberikan dampak kerugian sebab kebutuhan pakan pada ayam broiler 70% dari nilai produksi industri peternakan. Pencegahan dan pengendalian coccidiosis selama ini di lingkungan peternakan menggunakan coccidiostat, namun dirasakan keefektifannya telah berkurang sebab anticoccidia dapat menimbulkan resistensi dan mengakibatkan akumulasi residu coccidiostat dalam karkas. Era globalisasi menyebabkan berbagai kemajuan metode yang dapat digunakan untuk mencegah coccidiosis, salah satunya menggunakan vaksinasi. Vaksinasi broiler untuk coccidiosis memiliki keunggulan yakni: lebih aman, praktis, dan lebih ekonomis namun bergantung pada rute vaksinasi yang diberikan. Salah satu penelitian kegunaan vaksinasi terhadap coccidiosis diteliti oleh Rashid et al (2012) mengenai perbandingan keefektifan vaksin isolat lokal Formalin inactivated Eimeria tenella (FEV-48) dengan vaksin Irradiated Eimeria tenella (IEV) terhadap ayam broiler yang telah diinfeksi Eimeria tenella. Pembuatan vaksin FEV-48 menggunakan ookista Eimeria tenella yang telah disporulasikan kemudian dicampur dengan formalin 3% selama 48 jam, kemudian dilakukan pencucian menggunakan larutan Phosphat Saline (PZ) dan

Adults of Gurltia paralysans were obtained from veins of the spinal cord subarachnoid space from three domestic cats presenting with chronic paraparesis/paraplegia from rural areas of southern Chile. Four adult nematodes were collected (2 males and 2 females) were recovered from cat 1, 14 adult nematodes (12 females and 2 males) from cat 2, and 12 nematodes (10 females and 2 males) were collected from cat 3. Parasite induced lesions that compromised subarachnoid vein microvasculature at the thoracic, lumbar, sacral spinal cord segments extending to conus medularis. Female nematodes measured 25 mm long (range = 25–30 mm) and 0.1 mm wide. Male measured a mean of 16 mm length (range = 13–18 mm) with a body diameter of 0.1 mm (range = 0.08–0.15 mm). The present study described structural features of G. paralysans, a rare parasite first reported in the 1930s, and provides additional reports on associated clinical and pathological findings in naturally infected domestic cats.

The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis1 and Hypoderma lineatum2 (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation.

Fecal analyses are becoming increasingly important for equine establishments as a means of parasite surveillance and detection of anthelmintic resistance. Although several studies have evaluated various egg counting techniques, little is known about the quantitative effects of pre-analytic factors such as collection and storage of fecal samples. This study evaluated the effects of storage temperature, storage time and airtight versus open-air storage on fecal egg counts. The experimental protocols were replicated in two study locations: Copenhagen, Denmark and Athens, Georgia, USA. In both locations, the experiment was repeated three times, and five repeated egg counts were performed at each time point of analysis. In experiment A, feces were collected rectally and stored airtight at freezer (−10 to −18 °C), refrigerator (4 °C), room (18–24 °C), or incubator (37–38 °C) temperatures. Egg counts were performed after 0, 6, 12, 24, 48, and 120 h of storage. In experiment B, feces were collected rectally and stored airtight or in the open air in the horse barn for up to 24 h. Egg counts were performed after 0, 3, 6, 12, and 24 h of storage. In experiment A at both locations, samples kept in the refrigerator showed no decline in egg counts, whereas storage in the freezer and incubator led to significantly declining egg numbers during the study. In contrast, storage at room temperature yielded marked differences between the two study locations: egg counts remained stable in the U.S. study, whereas the Danish study revealed a significant decline after 24 h. In experiment B, the Danish study showed no differences between airtight and open-air storage and no changes over time, while the U.S. study found a significant decline for open-air storage after 12 h. This difference was attributed to the different barn temperatures in the two studies. To our knowledge, this is the first study to evaluate the pre-analytic factors affecting egg counts in horses using an experimental protocol replicated in two contrasting geographic and climatic locations. Our results demonstrate that refrigeration is the best method for storage of fecal samples intended for egg count analysis, but that accurate results can be derived from fecal samples collected from the ground within 12 h of passage.

In order to better understand the enzootiology of trypanosomiasis caused by Trypanosoma evansi in the Brazilian Pantanal we examined domestic and wild mammals by microhematocrit centrifuge technique (MHCT), immunofluorescence antibody test (IFAT) and polymerase chain reaction (PCR). T. evansi infection was detected in all species sampled with exception of the sheep and the feral pig. High parasitemias were observed in capybaras (5/24), coatis (18/115), horses (31/321) and dogs (3/112). Among these species, only the capybaras did not develop anemia. Low parasitemias, only detected by PCR, were found in buffaloes (18/43), bovines (29/331), marsupials (1/4), small rodents (14/67), bats (7/18), and one armadillo (1/8). The highest prevalence of T. evansi infection was recorded in horses (73%), although no neurological signs in infected horses were observed. Diagnosis through standard parasitological tests and IFAT should be used with caution since they may overlook comprovedly infected horses. The relationship between ranch management and T. evansi infection in horse was investigated. The importance of other transmission mechanisms apart from the tabanids and reservoir hosts are discussed.

Fasciolosis, caused by the trematode Fasciola hepatica, is a zoonosis of economic importance in livestock that is emerging as a chronic disease in humans. The intermediate hosts are lymnaeid snails, in which diagnosis of infection is traditionally based on cercarial shedding, tissue sectioning and crushing.We developed a PCR assay for the sensitive and specific detection of F. hepatica in field-collected Lymnaea sp. snails. A primer pair was designed to amplify a 405 bp fragment of the cytochrome c oxidase subunit 1 gene of F. hepatica.The PCR assay showed a limit of detection of 10 pg of genomic F. hepatica DNA. No cross-reactions were observed with samples from other related trematode species or from the snail hosts Lymnaea columella and Lymnaea viatrix. DNA sequencing of the amplicon showed 100% homology with F. hepatica, and 75–89% homology with other trematodes on regions that did not include the entire set of primers.Two samples from Argentina were analysed. For snails in sample 1 (n = 240), identified as L. columella, the infection rate was 17.5 and 51.3% by direct examination and PCR, respectively. For snails in sample 2 (n = 34), identified as L. viatrix, the infection rate was 2.9 and 61.8% by direct examination and PCR, respectively. Differences in infection rates between these diagnosis methods were significant for both samples.Our PCR technique showed to be effective for detecting specific F. hepatica infections of low intensity in the intermediate host, and hence it could be used to study the epidemiological situation in a given area, as well as to assess host suitability for the parasite.

In this study, infrared thermography (IRT) was assessed as a means of detecting the changes in body temperature in rabbit coccidiosis, as well as determining IRT used in imaging and measuring the regional changes in skin temperature that occur in the rabbits during this infection. Rabbits were inoculated with 5 × 104 or 1 × 105 oocysts of Eimeria intestinalis, respectively. The group of control animals remained non-inoculated throughout the study. All measured temperatures (rectal, ocular surface and auricle pavilion temperatures) dropped significantly (P < 0.05) in rabbits infected with 1 × 105 oocysts. This group of rabbits also showed a medium negative correlation in rectal temperature (r = −0.49), ocular surface temperature (r = −0.49) and auricle pavilion temperature (r = −0.49) and a small negative correlation in live body weight gain (r = −0.22) depending on the number of days elapsed from the time of infection exposure.