Isolation of Babesia spp. from asymptomatic human beings

Babesiosis is a worldwide tick-borne protozoan disease of a variety of wild and domestic animals. For decades, various babesia strains were considered host-specific. However, babesiosis has been diagnosed as the cause of acute illness in 13 human beings in recent years, three of whom died. Studies of babesiosis in domestic animals indicate that for every clinically demonstrable case, there are hundreds of cases of latent infection. In an effort to determine if human babesia infections might be present in Mexico, we selected as our testing ground an endemic rural area along the Gulf Coast where continuous epizootics of equine, bovine, ovine and canine babesiosis were known to occur. Of the 101 individuals examined serologically by the indirect hemagglutination (IHA) test using Babesia canis as antigen, 38 reacted at titers from 1 : 10 to 1 : 80. Blood from these reactors was injected into splenectomized hamsters. Hamsters inoculated with blood from three of the individuals showed babesia in their peripheral blood. The growth of the organism was established by subpassages into additional hamsters. The persons from whom the organisms were isolated were asymptomatic for babesiosis. None of the 29 individuals residing in Mexico City, which represented the urban population in the survey, reacted in the IHA test. This is the first documented finding of the latent form of human babesiosis.     

Performance of a Johne's disease enzyme-linked immunosorbent assay adapted for milk samples from goats.

Antibody detection-based tests for paratuberculosis offer speed and economy, 2 diagnostic test attributes important to animal industries with narrow profit margins. Application of such tests to individual milk samples instead of serum samples can further improve testing efficiency and decrease testing cost. Accuracy of a commercial bovine paratuberculosis enzyme-linked immunosorbent assay (ELISA) adapted for use on goat serum and milk samples was determined. Fecal, blood, and milk samples were collected from 159 goats belonging to 2 Wisconsin goat herds with a prior history of paratuberculosis and 1 herd of 50 goats from a paratuberculosis-free Wisconsin herd. Fecal samples were cultured using the modified BACTEC 12B media. Sera were tested according to the manufacturer's instructions for bovine samples. Milk samples were centrifuged and mixed with the ELISA kit's Mycobacterium phlei-containing diluent at a ratio of 1:2. Using fecal culture as the "gold standard," the sensitivity of the ELISA on goat serum was 64% and the sensitivity of the ELISA on goat milk was 48%. The milk ELISA had higher agreement with fecal culture results (kappa = 0.525) than the serum ELISA (kappa = 0.425). ELISA specificity was 100% on both serum and milk. Regression analysis also showed good correlation between serum and milk S/P values (r2 = 0.67). Although less sensitive, the ELISA on goat milk samples appears to offer a useful, low-cost alternative for detection of goats with paratuberculosis that have progressed to the stage of shedding M. paratuberculosis in their feces.     

A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA.     

Effects of estradiol 17 beta implants on hematologic values and the chemiluminescence response of neutrophils of steers.

The effects of subcutaneous administration of a commercially available estradiol 17 beta implant on hematologic values and the chemiluminescence response of neutrophils were evaluated in 14 steers. Chemiluminescence and hematologic values were measured in treated (n = 8) and nontreated (n = 6) steers on days -14, -7, and -1 prior to implantation. Estradiol 17B was implanted into the treated group of steers on day 0, and blood samples were obtained from all steers on days 1, 2, 3, 4, 8, 15, 22, 29, 36, 43, and 50. The concentration of estrogen in serum was significantly (P = 0.0120) higher following implantation. Chemiluminescence and hematologic indices were not significantly affected by either implant status or serum concentrations of estrogen. The results of this study suggested that the use of implants containing estradiol 17 beta for promotion of weight gain in steers will not result in alterations of hematologic values or the neutrophil respiratory burst.     

Computed tomographic anatomy of the head of the loggerhead sea turtle (Caretta caretta)

The heads of three loggerhead sea turtles were disarticulated and imaged immediately to minimize postmortem changes and then frozen and sectioned. For computed tomography (CT) imaging, the heads were positioned in ventral recumbency. Transverse CT images with soft-tissue window were obtained from the olfactory sac region to the temporomandibular joint region. After CT imaging, the heads were sectioned and the gross sections were compared to CT images, to assist in the accurate identification of the anatomic structures. Different clinically relevant anatomic structures were identified and labelled in two series of photographs (CT images and anatomic cross-sections). CT images provided good differentiation between the bones and the soft tissues of the head. The information presented in this paper should serve as an initial reference to evaluate CT images of the head of the loggerhead sea turtle and to assist in the interpretation of lesions of this region.     

Influence of cover crops on potential nitrogen availability to succeeding crops in a Southern Piedmont soil

Winter cover crops are essential in conservation tillage systems to protect soils from erosion and for improving soil productivity. Black oat (Avena strigosa Schreb) and oilseed radish (Raphanus sativus L.) could be useful cover crops in the southeastern USA, but successful adoption requires understanding their influence on N availability in conservation tillage systems. Black oat and oilseed radish were compared to crimson clover (Trifolium incarnatum L.) and rye (Secale cereale L.) for biomass production and effects on N mineralization during the summer crop growing season from fall 1998 through summer 2002 near Watkinsville, GA. Rye produced 40 to 60% more biomass, although N contents were less than the other cover crops. Oilseed radish and black oat N contents were similar to crimson clover. Black oat, oilseed radish, and crimson clover C/N ratios were less than 30, whereas rye averaged 39. Amount of N mineralized in 90 days (N _min90) measured with in situ soil cores was 1.3 to 2.2 times greater following black oat, crimson clover, and oilseed radish than following rye. No differences in N _min90 were found between black oats, crimson clover, and oilseed radish in 1999 and 2000. The amount of potentially mineralizable N (N ₀) was not different due to cover crop, but was 1.5 times greater in 2000 and 2002 than in 1999. The rate of N mineralization (k) was 20 to 50% slower following rye than the other three cover crops. Black oat and oilseed radish biomass production and soil N mineralization dynamics were more similar to crimson clover than to rye, which indicates that they could be used as cover crops in the southeast without significant changes in N recommendations for most crops.

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Anaplasma marginale: lack of cross-protection between strains that share MSP1a variable region and MSP4

In Mexico, there are no commercial alternatives for the immunoprophylaxis of bovine Anaplasmosis, a disease responsible for great economic losses. Blood derived Anaplasma marginale used for immunizing susceptible cattle has shown promising results for homologous protection and controversial results against unrelated strains. The present study examined, under controlled conditions, the cross-protective potential of an immunogen composed of blood derived A. marginale of three strains against challenge with strains not included in the immunogens. Groups 1 and 2 were immunized with blood derived Anaplasma from strains Mexico, Morelos and Yucatan, group 4 with strains Morelos, Veracruz and Yucatan, two more groups (2 and 5) of equal conditions were inoculated with an adjuvant alone. Groups 1, 4 and 5 were challenged with Mexico strain; groups 2 and 3 were challenge-inoculated with strain Veracruz; groups 3 and 5 with strains Veracruz and Mexico as controls. Only animals in group 1, immunized and challenged with strain Mexico showed adequate protection. Both groups challenged with strains not included in the immunogens developed poor protection, while all the controls had to be treated to prevent death.     

Piroplasma infection in dogs in northern Spain

Babesiosis is a world-wide zoonosis caused by tick-borne hematozoan parasites of the genus Babesia. Canine Babesidae have historically been classified as "large Babesia" (Babesia canis) and "small Babesia" (Babesia gibsoni) based on the size of their intraerythrocytic forms. Genetic sequencing technology using the polymerase chain reaction (PCR) has allowed further subdivision. B. gibsoni has three strains: "Asia", "California" and a recently identified small babesial-like parasite, Theileria annae. This newly recognised piroplasm appears to be hyperendemic in northwest Spain. In order to provide some insight into the situation, all the cases diagnosed in our laboratory (NW of Spain) during 2003 were evaluated. Our study (62 samples) shows the existence of a piroplasm morphologically different from B. canis and similar to B. gibsoni, which is genetically related to T. annae. Severe regenerative anemia and thrombocytopenia are almost constant characteristics of infection with T. annae in dogs. In many cases azotemia is found. Abnormally high serum concentrations of urea and creatinin, together with elevated concentrations of inorganic phosphorus, hypoalbuminemia, hypercholesterolemia, proteinuria, high protein/creatinin and presence of hyaline and granular casts in the microscopic examination of urine sediment suggest a glomerular component to the disease. We conclude that observational research and clinical trials should be conducted in order to improve our understanding of this emerging disease in order to provide some insight into the best therapeutic practices.     

Is the decline of birds and amphibians in a steppe lake of northern Patagonia a consequence of limnological changes following fish introduction?

• 1. Laguna Blanca, in Laguna Blanca National Park, is a lake in Patagonia which has been designated as a Ramsar site since 1971 because of bird diversity and abundance and importance for nesting, particularly for the black neck swan, Cygnus melanocoryphus. It is also valued for its populations of endemic amphibians, Atelognathus patagonicus and Atelognathus praebasalticus.
• 2. Avian and amphibian populations have decreased dramatically in recent years. Percichthys colhuapiensis, Percichthys trucha (Pisces, Percichthyidae), Salmo trutta and Oncorhynchus mykiss (Pisces, Salmonidae) were introduced into Laguna Blanca in 1965. Since 1986, no Atelognathus frogs have been found. The abundance of swans and coots, which are strongly associated with macrophytes for nesting and feeding, has diminished drastically, whereas piscivorous birds have increased.
• 3. The fishless condition of some neighbouring small lakes with abundant pond weeds, aquatic birds and endemic amphibians, was assessed in order to compare the physical and chemical characteristics and the quantitative composition of the benthos among lakes. Fish presence at Laguna Blanca and its absence at El Burro, Antiñir and Jabón lakes, were confirmed. Compared with previous results, it seems that the abundance of Amphipoda (Hyalella), Copepoda and Cladocera at Laguna Blanca has decreased, while Acari has increased and Notostraca has disappeared. Water transparency has diminished in Laguna Blanca and now is lower than that of fishless lakes.
• 4. P. colhuapiensis were captured only in Laguna Blanca, with the highest captures in the littoral zone. The population shows rapid individual growth in the early years and an absence of fish older than 6 years. The preponderance of benthos and the presence of macrophytes in the gut contents of adult P. colhuapiensis appear to indicate that they are important consumers of these resources.
• 5. This paper concludes that fish introduction in Laguna Blanca led to a complex trophic cascade effect (fish predation on tadpoles, fish competition for avian food, bottom disturbance, zooplankton reduction) producing deleterious effects on the amphibian and bird populations.

Copyright © 2006 John Wiley & Sons, Ltd.