Efficacy of the immunoblot assay for cysticercosis in pigs and modulated expression of distinct IgM/IgG activities to Taenia solium antigens in experimental infections

A recently invented immunoblot assay for human cysticercosis was evaluated for efficacy in pigs. The test population consists of 45 pigs with parasitologically confirmed cysticercosis, 47 with heterologous infections, 45 SPF or concrete raised control animals. With this group of 137 animals the test performance was 100% sensitive and 100% specific. The antigen-specific responses of immunoglobulin A (IgA), IgG and IgM in four pigs infected with Taenia solium eggs derived from a human were quantified by immunoblot. Antigen-specific activities were observed as early as 1 week postinfection. The first antigen-specific isotypic response was IgM antibodies directed against a glycoprotein at 97 KD (GP97). This activity generally disappeared between the sixth and ninth week postinfection. Between Weeks 5 and 8, IgG activity rose as IgM activity fell. The IgG activity, however, was directed mostly towards GP50 and GP42 antigens. If the same response occurs in people with cysticercosis, identifying specific isotype activity may help to distinguish new infection from old.     

The impact of zoonotic diseases transmitted by pets on human health and the economy

In the context of all causes of human morbidity and mortality, or even within the context of all infectious diseases affecting the public health, pet-associated zoonotic infections are of moderate importance. The data documented in Table 1, however, indicate that they do exact significant human health and economic costs. If complete data were available for all of the infections shown in Table 1, the costs would be considerably higher. Moreover, most of these diseases are preventable through educating the public, particularly pet owners, of the zoonotic potential of these diseases, so that they may take precautions to minimize the risks leading to infection. These measures include appropriate health care of pets to eliminate infectious agents, reducing the number of uncontrolled, ownerless pets as well as unwanted or poorly supervised pets, preventing pets from soiling public places with their feces, excluding animals from areas where children play, enforcing leash laws, and promoting responsible pet ownership. Veterinarians, physicians, and public health agencies can aid in these efforts; ultimately, however, the responsibility lies with the pet owner.     

Restoring Perennial Grasses in Medusahead Habitat: Role of Tilling,Fire, Herbicides,and Seeding Rate

Restoring arid regions degraded by invasive annual grasses to native perennial grasses is a critical conservation goal. Targeting site availability, species availability, and species performance is a key strategy for reducing invasive annual grass cover while simultaneously increasing the abundance of seeded native perennial grasses. However, the potential for establishing successful seedings is still highly variable in rangeland ecosystems, likely because of variable year-to-year weather. In this study, we evaluated the independent and combined inputs of tilling, burning, applying imazapic herbicide, and varying seeding rates on existing species and seeded native perennial grass performance from 2008 to 2012 in a southwestern Idaho rangeland ecosystem. We found that combining tilling, fire, and herbicides produced the lowest annual grass cover. The combination of fire and herbicides yielded the highest seeded species density in the hydrologic year (HY) (October ? September) 2010, especially at higher than minimum recommended seeding rates. Although the independent and combined effects of fire and herbicides directly affected the growth of resident species, they failed to affect seeded species cover except in HY 2010, when weather was favorable for seedling growth. Specifically, low winter temperature variability (few freeze-thaw cycles) followed by high growing season precipitation in HY 2010 yielded 14 × more seeded perennial grasses than any other seeding year, even though total annual precipitation amounts did not greatly vary between 2009 and 2012. Collectively, these findings suggest that tilling, applying prescribed fire, and herbicides before seeding at least 5 × the minimum recommended seeding rate should directly reduce resident annual grass abundance and likely yield high densities of seeded species in annual grass ? dominated ecosystems, but only during years of stable winter conditions followed by wet springs.     

Visceral leishmaniasis in a New York foxhound kennel

Although endemic throughout much of the world, autochthonous visceral leishmaniasis has been reported on only 3 previous occasions in North America. After diagnosis of visceral leishmaniasis in 4 foxhounds from a kennel in Dutchess County, New York (index kennel), serum and ethylenediamine-tetraacetic acid (EDTA)-anticoagulated blood were collected from the remaining 108 American or cross-bred foxhounds in the index kennel and from 30 Beagles and Basset Hounds that were periodically housed in the index kennel. Samples were analyzed for antibodies to or DNA of tickborne disease pathogens and Leishmania spp. Most dogs had antibodies to Rickettsia spp., Ehrlichia spp., Babesia spp., or some combination of these pathogens but not to Bartonella vinsonii (berkhoffi). However, DNA of rickettsial, ehrlichial, or babesial agents was detected in only 9 dogs. Visceral leishmaniasis was diagnosed in 46 of 112 (41%) foxhounds from the index kennel but was not diagnosed in any of the Beagles and Basset Hounds. A positive Leishmania status was defined by 1 or more of the following criteria: a Leishmania antibody titer > or = 1:64, positive Leishmania polymerase chain reaction (PCR), positive Leishmania culture, or identification of Leishmania amastigotes by cytology or histopathology. The species and zymodeme of Leishmania that infected the foxhounds was determined to be Leishmania infantum MON-1 by isoenzyme electrophoresis. Foxhounds that were > 18 months of age or that had traveled to the southeastern United States were more likely to be diagnosed with visceral leishmaniasis. Transmission of Leishmania spp. in kennel outbreaks may involve exposure to an insect vector, direct transmission, or vertical transmission.     

Prevalence of Taenia solium cysticercosis in swine from a community-based study in 21 villages of the Eastern Cape Province, South Africa

The pork tapeworm, Taenia solium, causative organism of porcine cysticercosis and human neurocysticercosis is known to occur in areas of South Africa including Eastern Cape Province but, despite increasing reports of its occurrence throughout the subregion, the prevalence is yet to be clearly established. The parasite presents a potentially serious agricultural problem and public health risk in endemic areas. The human populations considered to be at highest risk of infection with this zoonotic helminth are people living in rural areas most of whom earn their livelihood wholly or partially through livestock rearing. Here we report on initial results of a community-based study of pigs owned by resource-poor, emerging pig producers from 21 villages in the Eastern Cape Province. Lingual examination (tongue palpation) in live pigs, two enzyme-linked immunosorbent assays (ELISAs), which detect parasite antigen (B158/B60 Ag-ELISA and HP10 Ag-ELISA) and an enzyme immunotransfer blot (EITB) assay, which detects antiparasite antibody, were used to verify endemicity and estimate apparent prevalence. In the absence of a gold standard true prevalence was obtained, using a Bayesian approach, with a model that uses both available data and prior information. Results indicate that the parasite is indeed present in the study villages and that true prevalence was 64.6%. The apparent prevalences as measured by each of the four tests were: 11.9% for lingual examination, 54.8% for B158/B60 Ag-ELISA, 40.6% for HP10 Ag-ELISA and 33.3% for EITB. This base-line knowledge of the prevalence of T. solium in pigs provides information essential to the design and monitoring of sustainable and appropriate interventions for cysticercosis prevention and control.     

Transmission of visceral leishmaniasis through blood transfusions from infected English foxhounds to anemic dogs.

OBJECTIVE: To conduct serologic surveillance for Leishmania spp in English foxhounds from a kennel, as well as recipients of blood from these dogs, and determine whether L infantum organisms could be transmitted via blood transfusion. DESIGN: Serologic prevalence survey. ANIMALS: 120 English foxhounds and 51 dogs of various breeds receiving blood from these donors. PROCEDURE: Foxhound blood donors, foxhound nondonors, and nonfoxhound blood recipient dogs were evaluated serologically for Leishmania spp by indirect fluorescent antibody testing. Dogs that received packed RBC (PRBC) transfusions from foxhound donors from mid-1996 through mid-2000 were identified. Furthermore, dogs were serologically evaluated if they had received fresh frozen plasma (FFP) transfusions in 1999 and 2000 from seropositive foxhound blood donors. RESULTS: Thirty percent of the English Foxhounds were seropositive for Leishmania spp (titer > or = 1:16), although the degree of seropositivity varied considerably during the period. Furthermore, 57 foxhounds had been used as donors from 1996 to 2000, and 342 units of PRBC had been transfused to at least 227 patients. All 25 dogs screened that received PRBC from seronegative foxhound donors tested negative, whereas 3 of 7 dogs that received PRBC from seropositive donors tested positive. All 9 dogs that received FFP from seropositive foxhound donors remained seronegative. CONCLUSIONS AND CLINICAL RELEVANCE: To our knowledge, this report documents the first transmission of Leishmania spp by blood transfusion. The use of foxhounds as blood donors may not be advisable in North America.     

Improvements in the serodiagnosis of helminthic zoonoses

Remarkable progress has been achieved in developing improved serodiagnostic assays for a group of diseases for which other diagnostic methods are often lacking. Toxocariasis, trichinellosis, dirofilariasis, Taenia solium cysticercosis and the cystic and alveolar forms of hydatid disease are occult infections in humans and sometimes in lower animal hosts. Although Strongyloides stercoralis achieves patency in humans, parasitologic diagnosis is often very difficult. Efforts to develop reliable immunodiagnostic methods have spanned several decades but progress had been slow until recently. The complexity and nonspecificity of helminth antigens were major problems which prevented the full realization of the benefits of the highly sensitive assay systems now available. Modern immunologic methods including hybridoma technology, immunoaffinity chromatography and immunoblotting, however, have yielded improved reagents and the means to characterize their nature and function. The outcome of this research has been more sensitive and specific serologic tests based on measurement of both circulating antigens and antibodies as well as improved understanding of the nature of host-parasite interactions. Although much remains to be done, many improved immunodiagnostic procedures are already being applied in clinical diagnosis, epidemiologic studies and control programs directed against the helminthic zoonoses.     

Efficacy of divided doses of GS-23654 against immature Echinococcus granulosus infections in dogs.

One hundred dogs were used to evaluate the efficacy of GS-23654 (4-nitro-4'-isothiocyano-diphenyl-ether) against immature Echinococcus granulosus. A 25% suspension of active ingredient was administered at dose rates of 100, 200, or 400 mg/kg of body weight on 1, 2, or 3 occasions. Anthelmintic efficacy was dosage dependent and increased with the number of times the dosage was repeated. At none of the treatment schedules tested were all worms eliminated from all dogs, although 92.6% of the expected number of worms were eliminated from dogs given 400 mg/kg on 3 occasions.     

Determination of caffeine, theobromine, and theophylline in standard reference material 2384, baking chocolate, using reversed-phase liquid chromatography

A rapid and selective isocratic reversed-phase liquid chromatographic method has been developed at the National Institute of Standards and Technology to simultaneously measure caffeine, theobromine, and theophylline in a food-matrix standard reference material (SRM) 2384, Baking Chocolate. The method uses isocratic elution with a mobile phase composition (volume fractions) of 10% acetronitrile/90% water (pH adjusted to 2.5 using acetic acid) at a flow rate of 1.5 mL/min with ultraviolet absorbance detection (274 nm). Total elution time for these analytes is less than 15 min. Concentration levels of caffeine, theobromine, and theophylline were measured in single 1-g samples taken from each of eight bars of chocolate over an eight-day period. Samples were defatted with hexane, and beta-hydroxyethyltheophylline was added as the internal standard. The repeatability for the caffeine, theobromine, and theophylline measurements was 5.1, 2.3, and 1.9%, respectively. The limit of quantitation for all analytes was <100 ng/mL. The measurements from this method were used in the value-assignment of caffeine, theobromine, and theophylline in SRM 2384.