World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) second edition: guidelines for evaluating the efficacy of parasiticides for the treatment, prevention and control of flea and tick infestations on dogs and cats.

These second edition guidelines, updated from the 2007 version (Marchiondo et al., 2007), are intended to assist the planning and conduct of laboratory and clinical studies to assess the efficacy of ectoparasiticides applied to dogs or cats for the purpose of treating, preventing and controlling flea and tick infestations. Major revisions to this second edition include guidelines on the assessment of systemic flea and tick products, an update of the geographical distribution of the common fleas and ticks species on dogs and cats, determination of flea and tick efficacy based on geometric versus arithmetic means with respect to geographic regulatory agencies, modification of tick categorization in the assessment of efficacy, expanded guidelines on repellency and anti-feeding effects, enhanced practical field study guidance, and considerations on the ranges of flea and ticks for infestations in laboratory studies. The term ectoparasiticide includes insecticidal and acaricidal compounds, as well as insect growth regulators. The range of biological activities from animal treatment that are considered include: repellency and anti-feeding effects, knockdown, speed of kill, immediate and persistent lethal effects, and interference with egg fertility and subsequent development of off-host life cycle stages. Information is provided on the selection of animals, dose determination, dose confirmation and field studies, record keeping, interpretation of results and animal welfare. These guidelines are also intended to assist regulatory authorities involved in the approval and registration of new topical or systemic ectoparasiticides, and to facilitate the worldwide adoption of harmonized procedures.

Ivermectin and milbemycin oxime in experimental adult heartworm (Dirofilaria immitis) infection of dogs.

BACKGROUND: The US Food and Drug Administration reports an increase in the frequency of reports of lack of effectiveness claims for heartworm (HW) prevention products. HYPOTHESIS: At their labeled doses, single doses of commercially available HW prevention products are not completely effective against all field isolates of HW. ANIMALS: Forty-two HW-free dogs experimentally inoculated with a recent HW field isolate. METHODS: Placebo-controlled, blinded laboratory clinical trial. Dogs randomly allocated to 1 of 3 treatment groups with 14 dogs per group. Groups were untreated control or p.o. dosed with milbemycin oxime (MBO) or ivermectin (IVM). Dogs were inoculated with 50 HW third stage larvae 30 days before dosing and necropsy was performed on Day 123 after treatment to enumerate adult HW. RESULTS: Thirteen of 14 control dogs had adult HW detected at necropsy with a geometric mean worm count of 22.3. One HW was found in 1 dog in each of the MBO and IVM treatment groups. CONCLUSIONS AND CLINICAL IMPORTANCE: Two currently approved macrocyclic lactone HW preventives used at their labeled dose rates were <100% effective against a recent HW field isolate, supporting the hypothesis that the effectiveness of a single dose of these preventives can vary. This is important in guiding clients on expectations of product effectiveness.

Efficacy of a single whole-body spray treatment of spinosad, against Boophilus microplus (Acari: Ixodidae) on cattle.

The efficacy of a single whole-body spray of spinosad, a naturally derived control agent, applied at three concentrations was evaluated against cattle infested three separate times prior to treatment and at four weekly intervals following treatment with Boophilus microplus (Canestrini). At 0.0167% active ingredient (AI) both tick numbers (1894 ticks per calf) and index of fecundity (IF) of females (258.3) were no different than that of the control group. However, spinosad treatment at both 0.05 and 0.15% AI resulted in fewer ticks per calf (600 and 935, respectively) with lower IF values for females (43.4 and 38.4, respectively). The percent control of ticks on the animals at the time of treatment (acute efficacy) was dramatically lower at 0.0167% AI (21.4%) than at 0.05 (86.3%) and 0.15% AI (87.9%). Spinosad treatments appeared to be more effective against immature stages (nymphs and larvae) than against adult ticks that were on the animals at the time of treatment. The mean weight of females that survived to repletion was similar (322-348 mg) in all groups. By contrast, the mean weight of egg masses produced by females was highest in the control group (155 mg), whereas each increase in spinosad concentration resulted in a substantial decrease in egg mass weight, with the 0.15% AI group averaging only 73 mg. The hatch rate of eggs derived from females ranged from 93.4% in control females down to 53.9% hatch for females treated at 0.15% AI spinosad. The residual efficacy of spinosad at 0.0167% AI was poor even at 1 week following treatment, resulting in 101 ticks per calf and a level of control of only 66.4%. At 0.05% AI, protection against successful reinfestation was high at 1-week post-treatment where only five ticks per calf reached repletion, and control of the IF of these females was 99.3%. The 0.15% AI treatment provided almost complete protection against reinfestation for 2 weeks following treatment (< or =5 ticks per calf), and control of the IF of these ticks was >99.9%. Thus, the use of spinosad at US ports-of-entry would be unacceptable because of the critical necessity of achieving 100% control with a single treatment to prevent the reintroduction of ticks. However, it is likely ticks could be eradicated using spinosad in tick infested areas of the US if repeated (systematic) treatments were applied to cattle maintained on the premises.

Cerebral larva migrans in a raccoon (Procyon lotor).

During 1997, gross and histopathologic examinations were performed on an adult female raccoon (Procyon lotor) that was live-trapped in Corvallis, Oregon. Multifocal eosinophilic granulomas indicative of neural and visceral larva migrans were observed. However, within these granulomas, the presence of parasitic larva was seen only in the cerebrum. Morphologic characteristics indicated that the nematode was an ascarid larva. However, it was smaller than the larva of Baylisascaris sp. This appears to be the first documented case of cerebral larva migrans in a raccoon.

A retrospective histopathological survey of capillariasis in raccoons (Procyon lotor) from Oregon.

A 2-yr retrospective survey (1995-1998) of 63 adult raccoons (Procyon lotor) from Oregon, northwestern United States, was conducted by histopathological examination of various tissues. An overall prevalence of capillariasis was recorded in 52% of the raccoons. Highest prevalence of nematodes (29/30, 97.7%) was present in the anal sacs, where variable numbers of nematodes, their ova, and extensive diffuse inflammatory cellular infiltrate were evident. Lesser numbers of nematodes were also seen in the mucosa of the anterior alimentary tract (mouth 2/63, tongue 10/62, esophagus 2/63) and in the urinary bladder (7/63). However, at these sites, there was a paucity of inflammatory infiltrate. The findings of this study indicate that capillariasis of the anal sacs is commonly seen in free-ranging raccoons in Oregon, and the infection is essentially subclinical because only 1 case of anal sac impaction has been reported. Measurements of the nematode and their ova from the anal sacs indicate that it is most likely a new capillariid species, which to date has not been fully described in raccoons. It is speculated that the high prevalence of anal sac infection by a Capillaria sp. in Oregon raccoons may also be true for raccoons in other parts of North America and in parts of Europe and Asia where this species has been introduced.

Synthesis and endectocidal activity of novel 1-(arylsulfonyl)-1-[(trifluoromethyl)sulfonyl]methane derivatives.

We have recently synthesized a series of novel disulfonylmethane compounds that have shown anthelmintic and insecticidal (endectocidal) activity. Several analogues have shown activity against the internal nematode Haemonchus contortus. In sheep studies, these analogues have shown 100% control of this internal parasite at a 10 mg/kg rate. In vitro activity against the biting flies, Stomoxys calcitrans and Haematobia irritans, has been observed at rates as low as 25 and 2.3 ppm, respectively. Only marginal activity against the liver fluke Fasciola hepatica and Trichostrongylus colubriformis was seen. Respiratory control index values on rat liver mitochondria for this series suggested uncoupling of oxidative phosphorylation as a mechanism of action. Compound 1 is considered to be a promising agent for treatment of parasitized sheep.

Biochemical, biological, and genetic characterization of a sylvatic isolate of Trichinella.

Biological, biochemical, and genetic relationships of Trichinella isolates were assessed and compared from 3 species of Illinois fur-bearing mammals. Tongue muscle collected from 1987 through 1989 from 323 raccoons (Procyon lotor), 9 red fox (Vulpes fulva), and 1 coyote (Canis latrans) were digested and Trichinella muscle larval prevalences and mean intensities (larvae/g) determined. The prevalence and mean intensity of tongue muscle-stage larvae were 2.8% and 44.4% and 326 and 2 larvae/g for raccoon and red fox, respectively. The single coyote examined for muscle larvae was negative. Seven of 13 Trichinella isolates (5 raccoon, 2 red fox) were maintained and amplified in mice. Comparative analyses of DNA from larvae of these isolates and from other Trichinella isolates were performed by dot-blot hybridization using specific repetitive DNA probes. DNA from the 7 Illinois sylvatic isolates reacted with the Trichinella T5 isolate DNA probe pUPB-3.7 and did not cross-react with the Trichinella spiralis probe pBP-2. The sylvatic isolates of Trichinella were also analyzed using 27 allozymes. Their allozymic patterns were similar to Trichinella reference strain T5 from Pennsylvania, except for mannose phosphate isomerase. These results suggest that the Illinois wildlife isolates belong to the Trichinella T5 genotype according to the classification system established by the International Trichinella Reference Centre and as defined here by positive hybridization to the pUPB-3.7 probe. Results from Southern blot analyses using rRNA as a probe and allozyme patterns revealed some heterogeneity among geographical isolates of the T5 genotype.

Epidemiology of Ostertagia ostertagi in cow-calf herds in the southeastern USA.

Ostertagia ostertagi is commonly found in the brood cow and nursing calf in the southeastern USA, this information being derived from fecal egg counts, coproculture and necropsy results; however, clinical disease and large burdens of this parasite are rarely reported. Fecal egg counts in brood cows are routinely low and are generally reported to be 10 eggs per gram of feces (EPG) or less. Nematode egg counts in spring-born calves are also generally low prior to weaning; they increase steadily during the spring and summer and peak from late summer to fall weaning. That egg counts in spring-born calves are low for several months after birth is probably a reflection of minimal grazing activity. It appears that Ostertagia ostertagi may be of equal or less importance than other nematode genera for spring-born calves in the southeastern USA. The role that Ostertagia ostertagi plays in fall-born calves or in year-round calving herds has not been adequately investigated. Also, the role that the adult cow plays, with low egg counts and small Ostertagia ostertagi burdens, in contamination of pasture is not understood during either lactation or dry periods. Treatment of beef calves prior to, or at weaning can reduce contamination and transmission of gastrointestinal parasites on pastures which may be subsequently grazed by these or other weaned calves. Data on parasite population dynamics from tracer calf studies in cow-calf herds in the southeastern USA have identified peak periods of transmission and incidence of specific genera or stages. This information, in conjunction with routinely used cattle production and management practices such as time of calving, should provide means to more accurately define optimal timing for strategic parasite treatment programs and their overall effect on beef production in the southeastern USA.

First report of a Demodex sp. in raccoons (Procyon lotor).

Demodex spp. mites were seen in skin sections of 5 of 53 raccoons (Procyon lotor) necropsied on Parramore Island, Virginia (USA). In all infections, mites were present in the skin over the lower legs; in one raccoon they also were located in a follicle of a vibrissa. None of the raccoons had Demodex-related gross lesions. Histopathologically, minimal lesions were seen in the affected follicles. This is the first documentation of Demodex spp. in raccoons.

Helminth parasites of the raccoon (Procyon lotor) from north-central Arkansas.

Twenty-three species of helminths (4 trematodes, 2 cestodes, 14 nematodes, and 3 acanthocephalans) were found in the gastrointestinal tract, lungs, pancreas, tongue, urinary bladder, and subcutaneous tissues of 30 live-trapped or hunter-shot raccoons from north-central Arkansas between November 1989 and April 1990. Helminths were not detected in the brain, diaphragm, gallbladder, heart, liver, reproductive system, or trachea. Each raccoon examined was infected with 3 or more of the following helminths: Brachylaima virginiana, Eurytrema procyonis, Fibricola cratera, Pharyngostomoides procyonis, Atriotaenia procyonis, Mesocestoides spp., Arthrocephalus lotoris, Capillaria aerophila, Capillaria plica, Capillaria procyonis, Capillaria putorii, Crenosoma goblei, Cruzia americana, Dirofilaria tenuis, Dracunculus insignis, Enterobius sp., Gnathostoma procyonis, Molineus barbatus, Physaloptera rara, Trichinella spiralis, Centrorhynchus wardae, Macracanthorhynchus ingens, and Oligacanthorhynchus tortuosa. All helminths collected with the exception of D. insignis constitute new geographic distribution records. Occurrences of C. aerophila, C. wardae, and O. tortuosa are new host records. One nymphal pentastome, Porocephalus crotali, was found in the liver of 1 raccoon, constituting a new host record.

Prerequisites for oral immunization of free-ranging raccoons (Procyon lotor) with a recombinant rabies virus vaccine: study site ecology and bait system development.

A model baiting system suitable for the delivery of an oral rabies vaccine to free-ranging raccoons (Procyon lotor) was developed and tested on barrier islands in South Carolina (USA). Features of barrier island physiography and ecology were studied relative to selective bait deployment and site biosecurity. Capture-mark-recapture data were obtained from 228 raccoons. Raccoon density estimates, using a modified census assessment technique, were one raccoon per 1.8 to 2.7 ha. Mean (+/- SE) and range home area estimates of radio-collared raccoons were 84 (+/- 15.6) ha (27 to 176 ha) by a minimum convex polygon method and 138 (+/- 22.8) ha (43 to 241 ha), by a harmonic mean transformation method. Habitat utilization determinations of radio-collared raccoons were conducted to identify study areas to potentially maximize selectivity of bait towards raccoons and to reduce the absolute number of baits deployed. Island raccoons showed a habitat preference for maritime forest, maritime shrub and marsh areas. Additionally, there was no evidence of inter-island or mainland exchange of ear-tagged or radio-collared raccoons. A disease and mortality survey was conducted to identify baseline pathology and incidental lesions in the target raccoon population, prior to actual vaccination initiation. Thirty-eight percent of 30 clinically suspect raccoons sampled had intracytoplasmic eosinophilic inclusions diagnostic of canine distemper; no other lesions suggestive of viral etiologies were found. Serological surveys for raccoon poxvirus and rabies virus antibodies were negative. Antibody titers to canine adenovirus 1 and 2 indicated a moderate level of exposure (approximately 10 to 16%) in the raccoon population. Overall, 93 to 100% of placebo baits were consistently disturbed by 7 days post-bait deployment, and bait acceptance rates by raccoons ranged from 49 to 85%, by using a modular systems approach to select the optimum combination of bait attractant, biomarker, matrix, density, and distribution. These results suggest that a large proportion (up to 85%) of a free-ranging island raccoon population can be selectively and safely targeted, marked and monitored utilizing a proposed oral bait delivery system for recombinant or other rabies vaccines.

Notoedric acariasis in the porcupine (Erethizon dorsatum).

Six of 16 porcupines (Erethizon dorsatum) live-trapped in Sullivan County, Pennsylvania were infested with the mite Notoedres douglasi. Alopecia was evident in the ventral inguinal area and the infested skin was thickened, wrinkled and had multifocal areas of extensive hyperkeratosis. Microscopically the skin lesions consisted of extensive acanthosis and hyperkeratosis with variable cellular infiltrate in the dermis. Mites were seen in tunnels in the hyperkeratotic skin sections. This is the first report of notoedric acariasis in the porcupine.

Pancreatic duct hyperplasia in a raccoon (Procyon lotor) caused by Atriotaenia procyonis.

A raccoon (Procyon lotor) live-trapped in Philadelphia, Pennsylvania was heavily infected with the tapeworm Atriotaenia procyonis. Histopathologic examination of the duodenum revealed this tapeworm attached to the mucosa; the mucosa was mildly hyperplastic without a significant inflammatory infiltrate. This tapeworm was seen also within the proximal pancreatic duct and histopathologic lesions associated with its presence consisted of marked hyperplasia of mucosal cells, causing the formation of an extensive papillary projection into the duct lumen. Inflammatory cellular reaction was minimal, consisting primarily of lymphocytes, and was seen in the mucosa and submucosa of the duct. The presence of A. procyonis in the pancreas of this raccoon is considered to be an aberrant location for this parasite and has not been reported previously.

Lung lesions in an opossum (Didelphis virginiana) associated with Capillaria didelphis.

An opossum (Didelphis virginiana) live-trapped on Parramore Island, Virginia was infected with the lung parasite Capillaria didelphis. Lung lesions associated with this parasite were extensive and consisted of numerous areas with central necrosis deep within the parenchyma. Necrotic foci were present around airways and consisted of a mixture of inflammatory cells and cellular debris, adult nematodes and many bipolar eggs. This parasite causes severe verminous pneumonia in the opossum.

Lesions associated with pulmonary parasites in bobcats (Felis rufus) from Arkansas.

Two of five bobcats (Felis rufus) from southwestern Arkansas had natural pulmonary infections of Paragonimus kellicotti and Filaroides rostratus. Pairs of P. kellicotti were found in spherical cyst-like structures approximately 1 cm in diameter. Filaroides rostratus were seen as serpentine pale white areas on the pleural surface and also firmly embedded in fibrous capsules in the pulmonary parenchyma. Histologic lesions associated with the presence of these parasites consisted primarily of a verminous bronchitis. Clinical signs or compromised pulmonary function were not associated with these infections.

Sarcocystis kirkpatricki n. sp. (Apicomplexa: Sarcocystidae) in muscles of raccoons (Procyon lotor) from Illinois.

Sarcocysts of Sarcocystis kirkpatricki n. sp. (Apicomplexa: Sarcocystidae) are described from the skeletal and heart musculature of 66 (66%) of 100 raccoons (Procyon lotor) from Illinois. Histologic examination of muscle tissues from tongue, diaphragm, esophagus, and heart revealed that 61%, 47%, 32%, and 2%, respectively, contained sarcocysts of this species. Juvenile raccoons (less than 1 yr old) were more likely (P less than 0.01) to have sarcocysts in the tissues examined (52/60 or 87%) than were adults (14/40 or 35%). Histologically, sarcocysts in the 4 tissues were similar: the cyst wall was 2-3 microns thick, PAS negative, and had fine hairlike surface projections; interior septa were indistinct. Ultrastructurally, sarcocyst walls had short (mean = 2.8 microns), straight to sloping, villuslike projections. Longitudinal tubular filaments inside these projections extended from the tips to the base, where they terminated in a granular electron-dense layer of the primary cyst wall. Thin septa were within the sarcocysts. Feeding experiments utilizing dogs and cats as potential definitive hosts were negative.

Proposed field evaluation of a rabies recombinant vaccine for raccoons (Procyon lotor): site selection, target species characteristics, and placebo baiting trials.

Prior to a limited field application of an orally-administered vaccinia-rabies glycoprotein (V-RG) recombinant virus vaccine for wildlife, background data were obtained for the proposed site on Parramore Island, Virginia (USA). Mammalian target and nontarget species, potentially at risk for exposure to vaccine were inventoried. Placebo baiting trials with a fishmeal polymer bait resulted in high bait disturbance (88 to 100%), primarily by raccoons (Procyon lotor), with infrequent visitation and no evidence of bait consumption by deer (Odocoileus virginianus), small mammals or avian species. Definitive bait acceptance rates by raccoons (indicative of bait ingestion) were difficult to accurately determine based exclusively on premolar and vibrissae samples collected antemortem from live-trapped raccoons for tetracycline and rhodamine B biomarker analyses, respectively. Bait acceptance rate was more accurately determined during a pilot baiting trial conducted on North Island, South Carolina, when mandibles (postmortem samples) were examined for tetracycline incorporation. Parasitologic findings in raccoons on Parramore Island included Hepatozoan procyonis, Phagicola angrense and Physaloptera rara and a variety of incidental microscopic lesions, and provided baseline pathological data for comparison subsequent to V-RG vaccine application. A population density estimate of one raccoon/2.7 ha was calculated using mark-recapture data for comparison after vaccine deployment. Limited reproductive data, including estimates of pregnancy rates by palpation, the number of live kits/litter live-trapped with previously pregnant raccoons or observed in the dens of radio-collared raccoons, was gathered to assess the effect of proposed oral vaccination with V-RG vaccine. Home ranges were assessed by radio-telemetry of 15 raccoons; all radio-collared raccoons currently reside on Parramore Island.(ABSTRACT TRUNCATED AT 250 WORDS)

Dirofilaria immitis in a river otter (Lutra canadensis) from Louisiana.

A single adult male river otter (Lutra canadensis) from Louisiana was found naturally infected with Dirofilaria immitis. One adult male nematode was found in the heart; lesions attributable to the presence of the parasite were not found. This is the first report of D. immitis in the river otter in North America.