Multiyear Survey of Coccidia, Cryptosporidia, Microsporidia, Histomona, and Hematozoa in Wild Quail in the Rolling Plains Ecoregion of Texas and Oklahoma, USA.

We developed nested PCR protocols and performed a multiyear survey on the prevalence of several protozoan parasites in wild northern bobwhite (Colinus virginianus) and scaled quail (Callipepla squamata) in the Rolling Plains ecoregion of Texas and Oklahoma (i.e. fecal pellets, bird intestines and blood smears collected between 2010 and 2013). Coccidia, cryptosporidia, and microsporidia were detected in 46.2%, 11.7%, and 44.0% of the samples (n = 687), whereas histomona and hematozoa were undetected. Coccidia consisted of one major and two minor Eimeria species. Cryptosporidia were represented by a major unknown Cryptosporidium species and Cryptosporidium baileyi. Detected microsporidia species were highly diverse, in which only 11% were native avian parasites including Encephalitozoon hellem and Encephalitozoon cuniculi, whereas 33% were closely related to species from insects (e.g. Antonospora, Liebermannia, and Sporanauta). This survey suggests that coccidia infections are a significant risk factor in the health of wild quail while cryptosporidia and microsporidia may be much less significant than coccidiosis. In addition, the presence of E. hellem and E. cuniculi (known to cause opportunistic infections in humans) suggests that wild quail could serve as a reservoir for human microsporidian pathogens, and individuals with compromised or weakened immunity should probably take precautions while directly handling wild quail.

Genotypic and phenotypic evaluation for benzimidazole resistance or susceptibility in Haemonchus contortus isolates.

Haemonchus contortus isolates were evaluated for benzimidazole (BZ) resistance or susceptibility by allele-specific PCR based on ß-tubulin isotype 1 gene polymorphisms at the F167Y, E198A, and F200Y sites. Two isolates, one presumed susceptible from wild pronghorn antelope (PH) and one known to be resistant from goats (VM), were also assayed phenotypically for BZ resistance or susceptibility in the larval development assay (Drenchrite®). The BZ EC50 was 0.198 µM (intermediate between susceptible and weak resistant) for PH with critical well 5 (intermediate between susceptible and weak resistant) and 1.456 µM (intermediate weak resistant and resistant) for VM with critical well 8.5 (resistant). Genotypically, DNA extracted from pooled VM L3 larvae in the Drenchrite® wells with the highest BZ concentration was homozygous susceptible (SS) at the F167Y and E198A sites and homozygous resistant (RR) at the F200Y site by PCR, and sequence analysis bore this out. PH L3 larvae DNA from a control well (no BZ) was SS at all three sites by PCR, confirmed by sequence analysis. All single adult worm samples (N = 21) from PH, VM, Egypt goat (EG), and a Texas llama were SS at F167Y and E198A by PCR; however, only 3 PH worms and 1 EG worm were SS at F200Y. Three additional PH worms were RS and upon cloning two clones were identified as resistant by sequencing and two as susceptible. Clones from single adult worms VM, llama, and EG samples that were RR by PCR at F200Y were sequence verified as resistant. In this study, F200Y was the most frequently found genotypic marker for BZ resistance or susceptibility in the different Haemonchus isolates.

Detection and Management of Air Sac Trematodes (Szidatitrema Species) in Captive Multispecies Avian Exhibits.

From 2 exhibits in a zoological collection, 2 juvenile fairy bluebirds ( Irena puella ) and 1 adult blue-grey tanager (Thraupis episcopus) died within 3 months of one another. The cause of death was attributed to air sac trematodes, which were identified as Szidatitrema species based on morphology of adult trematodes and miracidia isolated from a snail intermediate host. To determine the extent of trematodiasis in the collection, individual exhibits within the same building as the original presenting cases were assessed, with birds representing 27 avian species from 9 orders. Sampling consisted of individual (n = 244) and pooled same-species group (n = 193) fecal examinations, and for some individuals, and tracheal swab (n = 106), resulting in a total of 543 samples. In addition, tracheal swabs were performed on 14 birds for comparative cytology, but no parasites were found. Flukes were positively identified in 4 tracheal swab samples (4%), 37 individual fecal samples (15%), and 52 of the group fecal samples (27%). When results of the swab method were compared with those of fecal examination, fecal testing was significantly associated (P < .001) with positive results. Based on these results, a screening process was instituted of 3- or 4-day combined fecal samples evaluated by both sedimentation and flotation techniques 2 weeks before outgoing shipments or intrazoo transfers of birds housed in exhibits known to have snail populations. Snail control methods also were initiated in all exhibits. Treatment with praziquantel was carried out on a case-by-case basis, and included oral, parenteral, and nebulized administration. Although control measures were expected to manage the infection and reduce distribution of the parasite to other collections, complete eradication of trematodes in the population is unlikely.

Morphological characterization of adult Fascioloides magna (Trematoda: Fasciolidae): first SEM report.

Five adult Fascioloides magna specimens were recovered from the livers of naturally infected calves from Texas, USA. Scanning electron microscopy was used to study the morphological characteristics of the trematodes. These mature flukes measured 35-100 mm in length by 15-25 mm in width and had an oval dorsoventrally flattened body, with no anterior cone. The tegument was armed with sharp spines. Around the oral and ventral suckers, some of the spines were small, with a sharp point, while others had serrated edges with 15-22 sharp points. The surface of the oral sucker was covered by an interesting pattern of tegument, small dome-shaped and ciliated papillae. The ventral sucker showed a smooth surface and two unknown spine-like structures. There were fewer spines at the base of the genital pore than on other parts of the anterior end of the worm. At the anterior end of the ventral side, well-developed spines were observed, while at the posterior end of the ventral side, the spines were small, mostly with one or three points and blunted edges. At the posterior end of the dorsal side, the spines became progressively fewer, smaller, and shorter. Around the excretory pore, the tegument was folded, with no spines, and small groups of dome-shaped and ciliated papillae were present. The cirrus organ showed a smooth surface, with small pores on the dorsal side and small groups of tiny spines between the folds. The eggs measured 168 × 101 µm and had a protoplasmic appendage at the pole opposite the operculum. At the posterior end of the dorsal side, and toward the right, a pore with a very thin rim was present, which could be the terminus of Laurer's canal.

Evaluating the effects of finishing diet and feeding location on sheep performance, carcass characteristics, and internal parasites.

A 3 yr experiment was conducted to evaluate the influence of diet and feeding location on animal performance, carcass characteristics, whole blood counts, and internal parasite burden of lambs assigned to 1 of 4 treatments: 1) confinement fed 71% alfalfa, 18% barley pellet, 5% molasses, 0.013% Bovatec, 6.1% vitamin/mineral package diet (CALF), 2) confinement fed 60% barley, 26% alfalfa pellet, 4% molasses, 2.5% soybean-hi pro, 0.016% Bovatec, 7.4% vitamin/mineral package diet (CBAR), 3) field fed 71% alfalfa, 18% barley pellet, 5% molasses, 0.013% Bovatec, 6.1% vitamin/mineral package diet (FALF), and 4) field fed 60% barley, 26% alfalfa pellet, 4% molasses, 2.5% soybean-hi pro, 0.016% Bovatec, 7.4% vitamin/mineral package diet (FBAR). A year × location interaction was detected for ending body weight (BW), average daily gain (ADG), and dry matter intake (DMI); therefore results are presented by year. In all years, cost of gain and DMI were greater for CALF and FALF than for CBAR and FBAR feed treatments (p ≤ 0.03). In yr 2 and 3 field treatments had greater ending BW and ADG than confinement treatments. For all years, diet did not affect ending BW or ADG. In yr 1 dressing percent and rib eye area were greater for field finished lambs than confinement finished (p ≤ 0.02) and Warner-Bratzler shear force was greater for CALF and FALF (p = 0.03). In yr 2 lambs in FALF and FBAR treatments had greater leg scores and conformation than CALF and CBAR (p = 0.09). In yr 1, FALF had a greater small intestine total worm count than all other treatments. In yr 1, ending Trichostrongyle type egg counts were greater for FALF (p = 0.05). In yr 2, ending Nematodirus spp. egg counts were greater for FALF and lowest for CBAR (p < 0.01). Abomasum Teladorsagia circumcinta worm burden was greater in CALF than all other treatments (p = 0.07) in yr 2. While field finishing lambs with a grain- or forage-based diet we conclude that it is possible to produce a quality lamb product without adverse effects to animal performance, carcass quality or increasing parasite burdens.

Anthelmintic resistant Haemonchus contortus in a giraffe (Giraffa camelopardalis) in Florida.

A young male giraffe (Giraffa camelopardalis) recently acquired by the Lion Country Safari in Loxahatchee, Florida, was diagnosed and successfully treated for Haemonchus infection while in quarantine. Seven weeks after introduction into a group of resident giraffes, this giraffe presented with diarrhea. Fecal evaluation revealed an extremely high count of 16,700 eggs/g, with larval identification of the parasite as Haemonchus. A larval development assay showed resistance to the three classes of anthelmintics currently used to treat Haemonchus contortus: the benzimidazoles, imidazothiazoles, and macrocyclic lactones. The giraffe was treated with a combination of moxidectin topically and fenbendazole orally, and follow-up fecal examination 2 wk later showed a marked reduction in strongyle-type eggs. However, within 2 mo the giraffe had a packed cell volume of 22% and an eggs per gram count of 11,900. The animal was then treated with moxidectin topically and copper oxide wire particles orally and removed from the contaminated area. Because of the unusual host, molecular analysis of the parasite was employed, which confirmed the nematode as H. contortus. It is likely that the monthly rotational deworming schedule first implemented more than 5 yr earlier contributed to the development of multiple anthelmintic resistance in this H. contortus population. The proper use of anthelmintics and good pasture management are crucial to reducing the parasite burden in captive giraffe.

Gastrointestinal Nematodes, Diagnosis and Control.

Disease caused by nematodes in the gastrointestinal tract of cattle is primarily economic and the effect of treatment is cattle grow larger faster because of increased feed intake. The disease, control measures, and drugs used must be focused on different ages and environments. Different drugs should be used against the parasites most likely affected by the specific drug and administered how and when it will accomplish the best sustainable control. Management needs to ensure that at-risk animals are exposed to sufficient worms to stimulate their immunologic response but not overwhelm it.

Anthelmintic resistance and alternative control methods.

Anthelmintic resistance has been a problem almost from the first use of the drugs introduced during the past 50 years. Evaluation of anthelmintics on a specific farm is essential to determine which drugs may be used in controlling parasite numbers. Treatment of livestock during the season in which parasite transmission is unfavorable lessens pasture contamination. Targeting treatment places less pressure on susceptible worms, diluting the resistant parasite population. Pasture management and using safe pastures for animals at highest risk lower pasture exposure. Selection for individual animals resistant to the effects of parasites lessens the need for use of anthelmintics. Alternatives to anthelmintics include cryptic antigen vaccines, copper wires, and biologic compounds.

Treatment of external and internal parasites of cattle.

To control parasitic disease in cattle, one first has to determine what parasites he or she is trying to control--nuisance and biting flies in a milking parlor or ticks on range cattle? The determination may be based on geographic location, class of cattle, and their management. After determining what is there, then if the numbers are sufficient, justify treatment. Each livestock producer has a different array of parasites with which to deal, and in some circumstances, parasitic disease is below the economic threshold. Others may have occasional clinical disease associated with parasites, but most have situations in which the potential of economic loss is high enough to justify control.

Demodex musculi in the Skin of Transgenic Mice.

Although infestations by a number of Demodex mite species have been described in mice, the occurrence of Demodex musculi infestation was last reported by Hirst in 1917. This communication describes the occurrence of D. musculi infestation in two lines of transgenic mice and their F1-hybrid offspring. We first found the Demodex mite in mouse hair samples collected during efficacy screenings in an ongoing ectoparasite treatment trial for the fur mite Radfordia affinis. An investigation was undertaken to determine the extent of the Demodex infestation within the facility and the original source of the parasite. D. musculi was found in three of the four mouse genotypes present in the index room and in one of these genotypes in two other rooms. The mite was not found in sentinel mice, other strains, or stocks within the facility. The mites were more easily recovered from the immunodeficient B6,CBA-TgN(CD3E)26Cpt transgenic (Tg) and the hybrid double-Tg (B6,CBA-TgN(CD3E)26Cpt x B6,SENCARB-TgN(pk5prad1)7111Sprd)F1 mice than from the B6,SENCARB-TgN(pk5prad1)7111Sprd Tg mouse, which is believed to be immunocompetent despite its thymic abnormalities. Histopathologic examination showed D. musculi superficially in hair follicles but not in the preputial or clitoral gland or in serial sections of the head, eyelids, or ears, the locations favored by other mouse demodicids. Physical and microscopic examination revealed no dermatitis. The immune deficiency in the B6,CBA-TgN(CD3E)26Cpt mouse probably provided the permissive host conditions that contributed to the proliferation and subsequent detection of the Demodex. Preliminary transmission experiments conducted with other immunologic mutant mice and our sentinel strain demonstrated variation in mite transfer and in either detection or maintenance of infestation when na ve mice were housed with those carrying D. musculi. The original source of D. musculi was not conclusively identified, but this parasite appears to be of low pathogenicity in the examined genotypes.

Seroprevalence of Anaplasma marginale in Texas cattle.

To our knowledge the seroprevalence of Anaplasma marginale in Texas has not been reported. The objective of this study was to estimate the point seroprevalence and spatial distribution of Texas cattle persistently infected with A. marginale. This was a cross-sectional observational study examining serum collected from 12,000 adult cattle marketed in 23 selected Texas auction markets during the second week of July 2011. A random subset of those cattle comprising 1835 individuals was evaluated for persistent infection with A. marginale using a commercial cELISA for antibody detection. The pooled apparent seroprevalence for cattle tested at auction markets across the state was 15.02% (95% CI: 11.02-19.53%), with markets in the western portion of the state demonstrating prevalence ⇒ 30%. The winter tick, Dermacentor albipictus is involved in the biological transfer of A. marginale and is prevalent in west Texas. Producers in endemic and non-endemic areas should be encouraged to determine the infection status of replacement cattle in order to implement effective management strategies for the control bovine anaplasmosis.

A new eimeriid (Apicomplexa) species from endangered Attwater's prairie chickens (Tympanuchus cupido attwateri) in Texas.

The Attwater's prairie chicken (APC; Tympanuchus cupido attwateri Bendire, 1894) has been a federally listed endangered species since 1967. Several captive propagation programs consisting of small populations are being used to keep this species from extinction. Fecal samples were collected from APCs in April 2007 and again in August 2008 from 2 separate captive propagation facilities in Texas after clinical signs of coccidiosis were observed. One Eimeria species was observed (Eimeria attwateri), which we describe as a putative new species. Sporulated oocysts are ellipsoidal, 30.0 × 18.4 (27.4-31.3 × 16.0-22.4) µm. Oocysts have a smooth wall 0.7 µm thick and lack both a micropyle and oocyst residuum, but 1 ellipsoidal polar granule is present, 2.3 × 1.9 (2.1-2.4 × 1.7-2.0) µm. Sporocysts have a nipple-like Stieda body with a rounded opposite end and are 14.0 × 7.1 (10.2-16.8 × 6.0-9.2) µm. The sporocysts contain a sporocyst residuum usually consisting of 2-4 dispersed globules, and each sporozoite contains 2 large posterior spheroid refractile bodies 3.4 µm wide. Nucleotide sequence amplified from the 18S rDNA does not match any DNA sequence information for publicly available Eimeria species, and phylogenetic reconstructions place this species with other eimerians from Galliformes. The discovery of a potentially pathogenic species of Eimeria in captive APCs is of great importance, and managers should be aware of the potential devastating effect(s) this parasite could have on the APC conservation programs.

Application of cognitive engineering principles to the redesign of a dichotomous identification key for parasitology.

Dichotomous identification keys are used throughout biology for identification of plants, insects, and parasites. However, correct use of identification keys can be difficult as they are not usually intended for novice users who may not be familiar with the terminology used or with the morphology of the organism being identified. Therefore, we applied cognitive engineering principles to redesign a parasitology identification key for the Internet. We addressed issues of visual clutter and spatial distance by displaying a single question couplet at a time and by switching to the appropriate next couplet after the user made a choice. Our analysis of the original paper-based key versus the Web-based approach found that of 26 applicable cognitive engineering principles, the paper key did not meet 4 (15%) and partially met 11 (42%). In contrast, the redesigned key met 100% of 32 applicable cognitive engineering principles.