An initial coprological survey of parasitic fauna in the wild Amur leopard (Panthera pardus orientalis)

The Amur leopard, one of nine recently recognized subspecies of leopard, is still the most threatened by a stochastic procession of extinction. Evaluation of the potential danger to the conservation of the Amur leopard originating from disease urgently needs to be studied. Unfortunately, research on the potential risk to Amur leopards caused by disease is rare. In terms of parasitic diseases that affect this species, even basic data for parasitic fauna are absent. The aim of this study is to acquire this knowledge to improve the general understanding of Amur leopard parasites. Seven parasite species, including 3 nematodes (Toxocara cati, a capillarid‐type parasite, and a Metastrongyloidea‐type parasite), 2 cestodes (Spirometra sp. and Taenia sp.), 1 trematode (Paragonimus sp.), and 1 protozoan (Cystoisospora felis), were found in this research. Toxocara cati occurred most frequently, followed by Spirometra sp.     

Detection of Myxobolus cerebralis in Rainbow Trout and Oligochaete Tissues by Using a Nonradioactive in Situ Hybridization (ISH) Protocol

Abstract

A nonradioactive in situ hybridization (ISH) protocol was developed to detect Myxobolus cerebralis, the causative organism of whirling disease, in its primary host, rainbow trout Oncorhynchus mykiss, and in its alternate oligochaete host, Tubifex tubifex. A cocktail of three oligonucleotide primers (derived from the small subunit ribosomal DNA sequence) directed at target sequences of the parasite DNA was tailed at the 3′ end with digoxigenin-labeled deoxyuridine triphosphate (DIG-dUTP). Labeled probes were hybridized to parasite DNA present in deparaffinized tissue sections from infected trout and oligochaetes. The bound probes were visualized after modifications of existing ISH protocols. By using the new ISH procedure, the parasite was found in target tissues of subclinically and clinically infected fish and tubificid oligochaetes after exposures of these hosts to triactinomyxons and mature spores, respectively. The probe did not bind with salmonid tissues infected with two other myxosporean parasites, Ceratomyxa shasta or the PKX organism, or to a Myxobolus sp. infecting the cartilage of plain sculpin Myoxocephalus jaok. These initial results indicate that ISH is an effective and specific test for detecting Myxobolus cerebralis in its fish and oligochaete hosts.     

Polystomatidae (Monogenea) of southern African Anura: Polystoma channingi n.sp. parasitic in two closely related Cacosternum species

Polystoma channingi is described as a new species of polystomatid flatworm (Monogenea) parasitic in the urinary bladder of Cacosternum nanum in the Eastern Cape Province, South Africa. In a locality where C. nanum and C. boettgeri occur sympatrically the parasite has been found in both species. This finding of a Polystoma sp. in two closely related host species questions the strict host-specificity generally ascribed to the genus, but supports the hypothesis that host-specificity is determined by the oncomiracidium's ability to recognize the tadpole as a suitable host. This is the first polystome to be described from the genus Cacosternum and the ninth species of Polystoma from South Africa. The species is distinguished by its huge haptor and suckers relative to the body size. The haptor length/body length ratio of 0.38 is by far the largest for the known southern African polystomes. The prevalence of infection with P. channingi n.sp. was 25% for C. nanum and 40% for C. boettgeri, while the mean intensity of infections in adult frogs was 1.5 for C. boettgeri and 1.0 for C. nanum.     

Small Subunit rRNA Gene Comparisons of Four Actinosporean Species to Establish a Polymerase Chain Reaction Test for the Causative Agent of Proliferative Gill Disease in Channel Catfish

Abstract

Proliferative gill disease (PGD) causes high morbidity and mortality in cultured channel catfish Ictalurus punctatus. The presence of the myxozoan Aurantiactinomyxon ictaluri (class Actinosporea) is strongly associated with PGD. This parasite, shed as an actinospore from the aquatic oligochaete Dero digitata, infects channel catfish by an undetermined route. Several other actinosporeans have been identified that are shed from D. digitata isolated from catfish ponds, including those designated A. mississippiensis, Helioactinomyxon sp., and the actinospore stage of Henneguya exilis. By the use of multiple sequence alignment of polymerase chain reaction (PCR)-amplified small subunit ribosomal RNA (SSU rRNA) genes of A. ictaluri, A. mississippiensis, and H. exilis, we identified two variable regions. The largest variable region was PCR amplified, sequenced from the Helioactinomyxon sp., and used in addition to the other three sequences in multiple-sequence alignment comparison to develop PCR primers specific for A. ictaluri. This PCR specific for A. ictaluri produced 104-base-pair products from a plasmid clone containing the SSU rRNA gene of A. ictaluri, spore DNA of A. ictaluri, and DNA prepared from channel catfish gill and D. digitata infected with A. ictaluri. The PCR assay was able to detect as few as 100 copies of the cloned gene. There was no detectable product from the genomic DNA of H. exilis, A. mississippiensis, or Helioactinomyxon sp., specific pathogen-free channel catfish gill, and noninfected D. digitata. The PCR assay will be useful as a diagnostic tool for PGD in channel catfish and will aid in the elucidation of the life cycle of A. ictaluri.     

New report of parasite-fauna of the free-tailed bat (<Emphasis Type="Italic">Tadarida brasiliensis,</Emphasis> Geoffroy, 1824) in Chile

One hundred and ninety five specimens of free tailed bat (Tadarida brasiliensis) obtained from two regions of Chile were analized to determine parasite infection. From those specimens the endoparasites identified were: Trematoda: Acanthatrium lunatum, Limatuoides limatulus and Paralecithodendrium carlsbadensis; Cestoda: Vampirolepis sp.; Nematoda: Nochtia pilosus and Anoplostrongylus paradoxus. The ectoparasites identified were Chiroptonyssus robustipes, Ewingnana inaequalis and Notoedres lasionycteris all of them are acari species. Even thought the bat specimens are from the same species on both survey sites, the results differ for each site when the parasite species identified are compared.     

Molecular characterisation of Cryptosporidium isolates from pet reptiles

In this study, we investigated the presence of Cryptosporidium in 171 faecal samples from reptiles commonly used as pet animals. These include lizards belonging to the genera Eublepharis, Pogona, Chlamydosaurus, Hemiteconyx, Teratoscincus, Tiliqua, Iguana, and Chamaeleo, snakes of the genera Lampropeltis, Elaphe, Python, Boa and Corallus, and tortoises belonging to the genera Testudo and Kinixys. Cryptosporidium oocysts were detected by immunofluorescence using a commercially available kit and cryptosporidial DNA by amplification of a polymorphic fragment of the 18S rDNA and the HSP70 locus.Cryptosporidium was detected in 38.6% and 25.1% of the samples analysed by immunofluorescence and PCR, respectively. Molecular characterisation of the isolates confirmed that C. serpentis and C. varanii (syn. C. saurophilum) are the main species involved in infection in pet reptiles but also showed the presence of C. parvum and C. muris, as well as other species or genotypes of this parasite including the Cryptosporidium mouse genotype and Cryptosporidium tortoise genotype previously described in reptiles. In addition, a Cryptosporidium sp. was isolated from a chameleon and a python.     

Freshwater nematodes from South Africa. 8. New and known species of Actinolaimoidea

Afractinolaimus Andrássy, 1970 is regarded as valid, its diagnosis slightly emended, and the following species transferred to it: Paractlnolaimus capensis, P. minor, P. zairensis and P. magaliesmontanus. The name of the type species thus becomes A. magaliesmontanus, since it has priority and is regarded as conspecific with A. noblei, the designated type. Additional biometrical data and distribution records are presented for A. magaliesmontanus. A. zairensis is reported from South Africa. Information is given for several populations of P. microdentatus. Two new species of Neoactinolaimus are described: N. brachydorus n. sp. is characterized by an exceptionally short and stout odontostyle, and N. barbieri n. sp. is distinguished from the closely related N. vaalensis by a thicker cuticle, more prominently set-off head, corrugated vestibule, longer oesophagus and several other minor morphological differences. Egtitus Thorne, 1967 is regarded as valid, and the following combination proposed: Egtitus zealandicus (Clark, 1963) n. comb, (synonym: Actinolaimus zealandicus).     

HELMINTH AND ARTHROPOD PARASITES OF THE DOMESTIC CAT IN PAPUA AND NEW GUINEA

A survey of the parasites found in 117 cats from the Port Moresby area of Papua and New Guinea is reported. The helminth species not previously recorded on the Australian mainland included Joyeuxiella pasquelei, Ancylostoma tubaeformae and Euparadistomum sp. The heart-worm, Dirofilaria immitis, is a relatively common parasite of aged cats in the Territory. Arthropods decorded included two indigenous tick species lxodes confusus and Haemaphysalis noveguineae.     

Identification and characterization of a <Emphasis Type="Italic">Streptomyces</Emphasis> sp. isolate exhibiting activity against multidrug-resistant coagulase-negative Staphylococci

The resistance of 220 coagulase-negative Staphylococci (CNS) (associated with animal disease) to 13 antibiotics were determined using the disk diffusion method. 35.9% of multidrug-resistant coagulase-negative Staphylococci (MR-CNS) exhibited resistance to five or more than five antibiotics; all of these bacteria were resistant to methicillin too. The new Streptomyces sp. ABRIINW111 was isolated from the Zagros Mountains Hamadan, Iran. The 16S rDNA sequence of the isolate indicated that it has 98% similarity to S. levis, but some mutations in the alpha and gamma regions of the 16S rDNA sequence emphasize the probability of the existence of a new species. Preliminary and secondary antibacterial screenings revealed that the isolate is active against gram negative and positive bacteria. The diethyl ether extracted metabolite of the Streptomyces sp. ABRIINW111 showed an effective antibacterial activity against MR-CNS. So the diethyl ether extract of the new Streptomyces sp. strain ABRIINW111 can inhibit the MR-CNS in vitro, and it can offer a new approach to treat MR-CNS infectious patients.     

New Host and Locality Records for Copepod and Isopod Parasites of Colombian Marine Fishes

Abstract

Two copepods and three isopods, representing five families of Crustacea, are noted from ten hosts, eight of these representing new host records. Cardiodectes rubosus (Copepoda: Pennellidae) is reported from the Atlantic Ocean for the first time. Excorallana tricornis (Isopoda: Corallanidae) may be a more important fish parasite than previously thought.     

Experimental in vitro transmission of Babesia sp. (EU1) by Ixodes ricinus

Babesia sp. (EU1), first characterized in 2003, has been implicated in human cases of babesiosis in Italy, Austria and Germany. It has been identified in roe deer and in its suspected tick vector, Ixodes ricinus, in several European countries. The aim of the present study was to validate the competence of I. ricinus as a vector of Babesia sp. (EU1) via experimental infections. For this purpose, a parasite strain isolated from roe deer was cloned in sheep erythrocytes. After experimental infections, parasite DNA was successfully amplified by PCR in both eggs and larvae originating from infected I. ricinus females and in the salivary glands of females exposed to Babesia sp. (EU1) as nymphs. We also demonstrate that infected females were able to transmit parasite DNA during a new blood meal. Together with previous epidemiological studies, these results validate I. ricinus as a competent vector for Babesia sp. (EU1).     

Helminths from three species of African chameleons

Three species of African chameleons (Chamaeleo gracilis, C. jacksoni and C. johnstoni) were examined for helminths. Two species of Cestoda (Oochoristica truncata and tetrathyridia of Mesocestoides sp.) and four species of Nematoda (Oswaldocruzia chamaeleonis, Parapharyngodon kenyaensis,Raillietnema chamaeleo, and Strongyluris elegans) were found. Four new host records are reported.     

Comparison of the susceptibility of brown trout (Salmo trutta) and four rainbow trout (Oncorhynchus mykiss) strains to the myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD)

Differences in susceptibility to the myxozoan parasite Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), between four strains of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were evaluated. Fish were exposed to water enzootic for the parasite in the field for 5 days and were subsequently transferred to the laboratory. Relative parasite load was determined after 2, 3 and 4 weeks post-exposure (wpe) by quantitative real-time PCR (qPCR) of kidney samples and number of parasite stages was determined in immunohistochemical stained sections of kidney, liver and spleen tissues. According to qPCR results, the highest amount of parasite DNA per equal amount of host tissue at all time points was measured in brown trout. Two of the rainbow trout strains showed lower relative parasite load than all other groups at the beginning of the experiment, but the parasite multiplied faster in these strains resulting in an equal level of relative parasite load for all rainbow trout strains at 4 wpe. A weak negative correlation of fish size and parasite load was detected. Only in samples of a few fish, single stages of T. bryosalmonae were found in sections stained by immunohistochemistry impeding quantitative evaluation of parasite numbers by this method. The results indicate a differential resistance to T. bryosalmonae between the rainbow trout strains investigated and between rainbow trout and brown trout.     

Some Copepod and Isopod Parasites of Bermuda Marine Fishes

Abstract

Three species are added to the depauperate parasite fauna of Bermuda: Caligus balistae, Lernanthropus talipes, and Alcirona insularis. The latter, normally a rare associate, seems to thrive on groupers (Serranidae) in captivity. An offshore record for Nerocila lanceolata on ocean triggerfish Canthidermis sufflamen contrasts with its known nearshore occurrence. A new host record is noted for each of four parasites occurring in Bermuda: Caligus balistae, A. insularis, Cymothoa oestrum, and N. lanceolata     

Susceptibility of Six Bacterial Pathogens of Channel Catfish to Six Antibiotics

Abstract

Two hundred eight bacterial isolates from diseased channel catfish Ictalurus punctatus were screened for susceptibility to apramycin, enrofloxacin, cephalothin, oxytetracycline, sulfadimethoxine-ormetoprim, and tilmicosin by the Kirby-Bauer disk diffusion method. Bacteria evaluated were Edwardsiella ictaluri, E. tarda, Aeromonas sp., A. hydrophila, A. sobria, and Pseudomonas sp. All bacteria were most susceptible to enrofloxacin (99.0%) and apramycin (97.6%), but only 86.5% were susceptible to sulfadimethoxine-ormetoprim, 84.1%o to oxytetracycline, and 75.5% to cephalothin. Edwardsiella ictaluri, E. tarda, A. hydrophila, and A. sobria were 100% susceptible to enrofloxacin. Aeromonas sp., E. ictaluri and E. tarda were 100% susceptible to apramycin. Resistance was detected in all bacterial species to cephalothin, oxytetracycline, sulfadimethoxine-ormetoprim, and tilmicosin. Testing E. ictaluri against sulfadimethoxine-ormetoprim gave larger zones of inhibition on Mueller-Hinton medium than on brain-heart infusion agar.     

Infecting Pacific Herring with Ichthyophonus sp. in the Laboratory

The protistan parasite Ichthyophonus sp. occurs in coastal populations of Pacific Herring Clupea pallasii throughout the northeast Pacific region, but the route(s) by which these planktivorous fish become infected is unknown. Several methods for establishing Ichthyophonus infections in laboratory challenges were examined. Infections were most effectively established after intraperitoneal (IP) injections with suspended parasite isolates from culture or after repeated feedings with infected fish tissues. Among groups that were offered the infected tissues, infection prevalence was greater after multiple feedings (65%) than after a single feeding (5%). Additionally, among groups that were exposed to parasite suspensions prepared from culture isolates, infection prevalence was greater after exposure by IP injection (74%) than after exposure via gastric intubation (12%); the flushing of parasite suspensions over the gills did not lead to infections in any of the experimental fish. Although the consumption of infected fish tissues is unlikely to be the primary route of Ichthyophonus sp. transmission in wild populations of Pacific Herring, this route may contribute to abnormally high infection prevalence in areas where juveniles have access to infected offal.

Received June 25, 2015; accepted September 12, 2015     

Mixed Metazoan and Bacterial Infection of the Gas Bladder of the Lined Seahorse—A Case Report

Abstract

Five wild-caught Lined Seahorses Hippocampus erectus from an aquarium system presented with altered buoyancy and distended upper trunks. Radiography of one specimen revealed a reduced air volume in the gas bladder. Pneumocystocentesis revealed a brown exudate of numerous leukocytes, parasite ova, and Gram- and acid-fast-positive bacilli under wet mounts and stains. Necropsies revealed enlarged, friable kidneys and distended gas bladders containing copious purulent exudate, necrotic tissue, and adult digeneans Dictysarca virens. Bacterial isolates from exudate cultures grown on Lowenstein–Jensen medium were identified as Gordonia sp. and Mycobacterium poriferae by high-performance liquid chromatography and 16S ribosomal DNA sequencing. Histopathology demonstrated a histiocytic response in kidney and gas bladder exudate, inflammation of the gas bladder wall, and infection of the gas bladder lumen with parasite ova and acid-fast-positive and Gomori's methenamine silver-positive bacilli. Praziquantel is prescribed for digenean infections but dissolves incompletely in seawater and is toxic to this host. Eradication of intermediate host vectors is a management option. Treatment of Gordonia infection has not been addressed in nonhuman animals, and there is no known effective treatment for Mycobacterium spp. infection in fishes. This is the first case report of digenean infection of the gas bladder in a syngnathid, Gordonia sp. infection in a nonhuman animal, and M. poriferae infection in a fish.

Received May 25, 2012; accepted October 12, 2012     

Anti-babesial activity of a potent peptide fragment derived from longicin of Haemaphysalis longicornis

Babesiosis is one of the most important tick-borne diseases affecting livestock that can cause major economic losses worldwide particularly in the tropics. Control relies on controlling both the protozoan parasite and the tick vector. Antiprotozoal drugs are most commonly used for treatment, but problems on emergence of resistant strains and food residues are encountered. Longicin, a defensin-like peptide identified from the hard tick, Haemapysalis longicornis, as well as one of its synthetic partial analogs (P4), were previously reported to exert antimicrobial, fungicidal, and parasiticidal activity. Both longicin and P4 showed babesiacidal activity, in vitro and in vivo. Here, peptide fragments of P4 were studied for in vitro activity against bovine Babesia parasites. One of the peptide fragments, antimicrobial peptide 1 (AMP1), reduced the parasitemia of Babesia bigemina. No peptide had significant effect on Babesia bovis. The sequence of AMP1 corresponded to the longicin sequence which is associated with antiparasitic activity. Although AMP1 caused reduction in parasitemia of B. bigemina, the difference in morphology of the parasite compared with the control group was not statistically significant. However, the percentage occurrence of piroplasms decreased, whereas the abnormal pycnotic form increased. The results demonstrated that this shorter peptide retained the anti-babesial activity of the parent peptide, exerting an antiparasitic effect against a bovine Babesia species. Therefore, this short peptide can be considered for chemical synthesis as an alternative therapeutic agent for babesiosis.