Dogs are definitive hosts of Neospora caninum

Dogs were investigated to determine if they are definitive hosts of Neospora caninum. Four dogs were fed N. caninum tissue cysts in infected mouse tissue, and two negative control dogs were fed uninfected mouse tissue. Dog faeces were examined daily for 30 days using a sucrose flotation technique. Three challenged dogs shed spherical to subspherical unsporulated oocysts, measuring 10 to 11 microns in diameter. Oocysts sporulated within 3 days and contained two sporocysts, each with four sporozoites. Outbred, inbred, and gamma-interferon knockout mice were inoculated with canine faecal extracts and monitored for evidence of neosporosis using a variety of morphologic, immunohistologic, serologic, and genetic analyses. Mice that received faeces from each dog observed to shed oocysts were demonstrated to have neosporosis by two or more techniques. One mouse was demonstrated to be infected with N. caninum by immunohistochemistry, ultrastructural analysis, and a species-specific PCR test. No evidence of neosporosis was observed in control animals. Based on this study, dogs are a definitive host of Neospora caninum.     

Oral Neospora caninum inoculation of neonatal calves

The bcl-2 protein plays an essential role in preventing cell death. Its activity is regulated through association with bcl-2 homologous and nonhomologous proteins and also by serine phosphorylation. We now report that bcl-2 can be proteolytically cleaved towards its N-terminus by a cysteine proteinase present in RL-7 lymphoma cell lysates, yielding a major product of apparent MW 20 kDa, different from the products of bcl-2 cleavage by HIV protease. Moreover, bcl-2 proteins mutated for Asp residues at positions 31 and 34 were efficiently cleaved by RL-7 cell lysates, indicating that this proteolytic activity is distinct from the caspase-3 that cleaves bcl-2 at Asp 34. This bcl-2 cleaving activity is inhibited by E-64 and is therefore distinct from the proteinases of the ICE/Ced-3 family (caspases), whereas reciprocally, ICE (caspase-1) is unable to cleave bcl-2. It is optimally active at pH 5, a feature distinguishing it from calpain, another non-ICE cysteine proteinase which has been associated with apoptosis. This novel bcl-2 cleaving protease, although constitutively present in RL-7 cells and resting peripheral blood lymphocytes (PBL) was upregulated following induction of apoptosis in RL-7 cells or mitogen activation in PBL. The N-terminus of bcl-2 which contains the BH4 domain that binds the kinase Raf-1 and the phosphatase calcineurin is essential for anti-apoptotic activity. Its cleavage might provide a novel post-translational mechanism for regulating bcl-2 function and could amplify ongoing programmed cell death.     

Neospora caninum identification in an aborted bovine fetus in Spain

Neospora caninum tachyzoites were identified in areas of the cerebrum with lesions of non-suppurative encephalitis in an aborted bovine fetus in Spain.     

Serological survey of antibody to Neospora caninum in Japanese dogs

Prevalence of antibody to Neospora caninum (NC) in Japanese dogs were examined. The antibody was positive in 15 of 48 dogs (31.3%) reared in the dairy farms that had case of the abortions due to NC infection or had the cattle seropositive to NC, whereas the prevalence was 7.1% (14 of 198 dogs) among the dogs kept in urban areas. In one dog breeder, all 17 Shetland sheepdogs older than 7 months were seropositive, and one pup was diagnosed as neosporosis 2 months before the first serological examination. The antibody titers of the dogs kept at this breeder were almost unchanged for 1.5 years. Serological evidence of the dogs in the dairy farms and urban areas indicates the transmission of NC between dogs and cattle. Also serological results of the dogs in one breeder may suggest the potential horizontal transmission among dogs.     

Prevalence of antibodies to Neospora caninum in different canid populations

A total of 1,554 dogs from 5 countries on 3 continents were tested for antibodies to Neospora caninum using an indirect fluorescent antibody test. In Australia, overall, 42/451 (9%, 95% confidence interval [CI] 6-12%) dogs were seropositive (Melbourne 11/207 [5%, 95% CI 2-9%]; Sydney 18/150 [12%, 95% CI 7-18%]; Perth 13/94 [14%, 95% CI 8-22%]). Antibodies to N. caninum were also detected in dogs in South America (Uruguay [20%, 95% CI 16-24%, n = 414]) and sub-Saharan Africa (Tanzania [22%, 95% CI 12-36%, n = 49]). In contrast, only 1 of 500 dogs tested from the Falkland Islands and none of 140 dogs from Kenya was seropositive. Of wild canids, 1/54 (2%, 95% CI 0-10%) British foxes and 15/169 (9%, 95% CI 5-14%) Australian dingoes had antibodies to N. caninum.     

Profiles of reproductive hormones associated with fertile and nonfertile inseminations of dairy cows

Differences in plasma levels of estrogen, progesterone, luteinizing hormone (LH), and urinary estrogens between fertile and nonfertile matings in dairy cows were studied. Samples were collected at 8-hour intervals prior to ovulation (Day 0), every day through Day 8, and on alternate days during Days 10-18. Every nonfertile case had hormonal levels outside the limits found in 10 of 11 fertile cases. Of 27 cases, 30 which involved lactating cows, 32 had a palpable corpus luteum following insemination, and 11 were fertile. Delayed estrus and a delayed preovulatory increase in LH following a decrease in progesterone to less than .75 ng/ml were consistently observed in nonfertile matings. These cases represented 75% of those with subnormal postovulatory levels of progesterone. Asynchronies among nonfertile cases compared with fertile cases involved progesterone, LH, estrogens, and urinary estradiol-17beta prior to ovulation. Examples illustrative of abnormal hormonal profiles included ovarian cysts and early and late regression of the corpus luteum.     

Identification of a major surface protein on Neospora caninum tachyzoites

Neospora caninum is a recently identified coccidian parasite that is closely related to Toxoplasma gondii. Molecules associated with the surface of N. caninum tachyzoites are likely to be involved in the process of adhesion and invasion of host cells. They probably also participate in the interaction of the parasite with the immune system, and they could play an important role in the pathogenesis of the parasite. To identify such surface molecules, we performed subcellular fractionation studies of isolated N. caninum tachyzoites. Employing the nonionic detergent Triton-X-114, we prepared a membrane fraction. Immunoblot analysis of this fraction using polyclonal antisera directed against tachyzoites of N. caninum and T. gondii resulted in the identification of a protein of approximately 43 kDa (Nc-p43). This molecule was present in two isolates of Neospora (Nc-1 and Liverpool) but was absent in Toxoplasma (RH-strain) tachyzoites. Further immunofluorescence and immunogold transmission electron microscopy (TEM) studies using affinity-purified anti-Nc-p43 antibodies demonstrated the presence of this molecule on the surface of N. caninum tachyzoites.     

Vaccination of mice with Neospora caninum: response to oral challenge with Toxoplasma gondii oocysts

Squalamine is a novel cationic steroid that possesses potent, broad spectrum, antimicrobial activity. Recent data suggests that squalamine or related compounds may be present and important in host resistance to infection in the urinary tract. Therefore, the role of the K54 capsule and the O4 specific antigen moiety of the lipopolysaccharide in protecting an extraintestinal isolate of Escherichia coli against the bactericidal activity of this novel antimicrobial compound was studied. The O4 specific antigen was important for protection against squalamine. Surprisingly, in contrast, the presence of the K54 antigen enhanced the bactericidal activity of squalamine. This is the first example, to our knowledge, in which an established virulence trait, the K54 capsule, may be detrimental to an infecting pathogen under certain circumstances.     

Neospora caninum causes abortions in a cattle herd in North Rhine Westphalia]

In a cattle herd in North Rhine-Westphalia a series of eight abortions occurred between 15th August and 14th October, 1996. Four of the aborted fetuses were histopathologically, virologically, bacteriologically and parasitologically examined. A multifocal necrotising encephalitis was observed in two fetuses. In addition, the liver of two fetuses and the placenta of one fetus contained infected foci. In three fetuses protozoan stages were detected which were identified as N. caninum by immunohistochemistry. A polymerase chain reaction conducted with histological material revealed N. caninum-DNA in the placenta of an aborted fetus. When seven cows that had aborted were tested for antibodies directed against N. caninum, six animals were positive by immunofluorescence and all seven by immunoblotting. The serological examination of the entire herd revealed a high herd seroprevalence of antibodies directed against N. caninum. Potential modes of infection with N. caninum in the herd are discussed, in particular the possibility of exposure to a point source (definitive host).     

Practical experiences with prostaglandins in heifers and dairy cows

This paper describes the extent to which utilisation of prostaglandins (PG) to achieve synchronisation of oestrus can influence certain economic results. Special attention is paid to the fertility status (FS) and calving index in dairy cows, where PG-treated groups are compared with untreated groups. With regard to these parameters a significant difference exists in favour of the PG treated group. In heifers the pregnancy rate after the first insemination has been determined in a PG treated group. The result compare very well with other published data. On large farms, where heat detection is poor, the routine use of PGs in the total herd is economically attractive. On those farms where the practitioner is involved in a fertility guidance system and where optimum heat detection takes place, the routine use of PGs in dairy cows would seem to offer advantages from a financial point of view.     

Experimental reproduction of bovine fetal Neospora infection and death with a bovine Neospora isolate

It is well established that alpha 2-adrenoceptor agonists have sedative and antinociceptive properties. In the current behavioral study we tried to find out if the alpha 2-adrenergic sedative and antinociceptive effects can be dissociated. We tested the hypothesis that alpha 2-adrenergic sedation is mediated by the locus coeruleus (LC) and antinociception by spinal alpha 2-adrenoceptors. Also, we addressed the possibility that intracerebral injection of an alpha 2-agonist might produce its antinociceptive effect by an action directly at the spinal cord. Medetomidine, an alpha 2-adrenergic agonist, or atipamezole, an alpha 2-adrenergic antagonist, were microinjected bilaterally into the LC through chronic cannulae in unanesthetized Han-Wistar rats. The effect on locomotor activity (/vigilance), tail-flick and hot-plate response, and on formalin-induced pain behavior was determined. Medetomidine microinjected into the LC (1-10 micrograms/cannula) produced dose-dependently hypolocomotion (/sedation), increase of response latencies in the hot-plate and the tail-flick tests, and a decrease in the formalin-induced pain behavior. Hypolocomotion (/sedation) was obtained at a lower medetomidine dose (1 microgram/cannula) than antinociception (3-10 micrograms/cannula). The lowest medetomidine dose used (1 microgram/cannula), which induced significant hypolocomotion (/sedation), produced either no antinociception (hot-plate and tail-flick tests) or even a slight hyperalgesia (formalin test). The hypolocomotion (/sedation) but not antinociception (tail-flick test) induced by systemic administration of medetomidine (100 micrograms/kg s.c.) could be reversed by atipamezole (10 micrograms/cannula) microinjected into the LC. Only a high systemic dose of atipamezole (1 mg/kg s.c.) reversed the antinociceptive effects of medetomidine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Association of abnormal uterine discharge with new intramammary infection in the early postpartum period in multiparous dairy cows

The association of abnormal uterine discharge with the development of intramammary infection (IMI) was studied in 62 multiparous Holstein cows during the nonlactating period and from lactation days 3 through 30. Duplicate milk samples were obtained from each mammary gland at approximately day 30 of the nonlactating period. Milk samples for bacteriologic culture also were obtained from each gland from all cows at the end of the previous lactation, at parturition, and on a minimum of 7 additional dates during the first 30 days of lactation. Beginning after parturition and continuing once weekly for 4 weeks, each cow was examined, using a vaginal speculum to visually estimate the quantity of abnormal uterine discharge in the vagina. Additionally, uterine swab specimens were obtained for aerobic bacteriologic culture. Cows were allotted to groups on the basis of the maximal amount of abnormal uterine discharge observed at any 1 of the 4 examinations. Cows in group 1 had normal discharge or < 30 ml of abnormal discharge; in group 2, > or = to 30 ml of abnormal discharge, observed only on examination by vaginal speculum; and in group 3, > or = to 30 ml of abnormal discharge visible externally. A difference was not detected in the development of new IMI in the nonlactating period between cows that subsequently developed uterine discharge and those that did not. Although significant differences were not found, a tendency for lactating cows with abnormal uterine discharge to be at increased risk for developing new IMI was observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Facility for exercising dairy cows

That dairy cows housed in confinement might benefit if they were exercised during the dry period prompted development of a facility for exercising dairy cows. Two methhods of the exerciser, which consist of a circular lane and a driving mechanism powered by a variable speed electric motor, are described. A speed of 3.5 k, per h provides a steady but comfortable pace for pregnant dairy cows and heifers. Many animals refused to go at speeds greater than 5.5 km per h.     

Prevalence of Neospora caninum and Toxoplasma gondii antibodies in sera from camels from Egypt

Sera from camels from Egypt were examined by the direct agglutination tests incorporating mercaptoethanol for antibodies to Neospora caninum and Toxoplasma gondii. Antibodies to N. caninum were found in 6 of 161 camels in titers of 1:40 (2 camels) and 1:80, 1:160, 1:640, and 1:1280 in 1 camel each, using N. caninum formalin preserved whole tachyzoites as antigen. Antibodies to T. gondii were found in 17.4% of 166 camels in titers of 1:25 (3 camels), 1:50 (18 camels). and > 1:500 (8 camels) using T. gondii tachyzoites. All 6 camels with N. caninum antibodies had no T. gondii antibodies in 1:4 dilution of serum, indicating specificity of the reaction. This is the first report of N. caninum prevalence in Egypt.     

Diagnosis of fatty liver in dairy cows

From 27 dairy cows with a mean milk yield of 6900 kg FCM (4% milk fat) per 305 day lactation period liver bioptates 2 weeks post partum (p.p.), milk samples 2, 4 weeks p.p., blood samples 0 (partus), 2, 4 week p.p., measurement of backfat thickness 2 weeks prior to calving, 0, 6, 17 weeks p.p. were taken and body weight and milk yield were determined. Fertility results and disorders appearance were recorded too. Total lipid and triglyceride content were analysed in liver tissue. Acetone concentration was determined in milk and 15 clinical-chemical parameters were elucidated in blood samples. Liver fat concentration shows a great variability from 3.9% to 24%. There is no strong reference value for the distinction between physiological and pathological liver fat concentration. Assessment as to whether increased liver fat levels in dairy cow are indicative of liver damage due to a pathological process should include detection of liver cell damage on the basis of plasma enzymes with closest possible specificity of liver. Glutamate-dehydrogenase (GLDH) is recommended. Liver fat content clearly could be defined exclusively from investigation of liver tissue rather than from analysis of blood or milk parameters. Measurement of backfat thickness provided useful information on the post partum lipolysis rate with a good correlation to liver fat (r to -0.72).     

Polymerase chain reaction approaches for the detection of Neospora caninum and Toxoplasma gondii

This review summarizes existing knowledge on the development and use of the polymerase chain reaction for the detection of DNA from Neospora and Toxoplasma. Several strategies which utilise the polymerase chain reaction for the diagnosis of toxoplasmosis in humans and livestock have been described and they principally target the B1 repetitive sequence, the P30 gene or ribosomal DNA. Experience has shown that the polymerase chain reaction has proven insufficiently robust to serve as a diagnostic test alone although when used in conjunction with other diagnostic techniques it does prove to be a useful aid. The marketing of a commercial polymerase chain reaction kit may well solve some of the inadequacies seen using "home made" polymerase chain reaction technology which are commonly used in diagnostic laboratories around the world. Recent progress on the development of polymerase chain reaction diagnostics for Neospora has been rapid and is discussed in detail.     

Experimental infection of dogs with the nasal mite Pneumonyssoides caninum

Cisplatin (CP) has been reported to activate murine macrophages to tumoricidal state, however, its mechanism of action is not known. In the present study it is reported that the production of: (a) interleukin-1 (IL-1); (b) tumor necrosis factor (TNF); (c) nitric oxide (NO); and (d) macrophage-mediated cytotoxicity by cisplatin-treated bone marrow-derived macrophages were inhibited by PKC inhibitors H-7 and chelerythrine chloride. Also, it was observed that treatment of macrophages with CP resulted in the translocation of PKC from the cytosol to the membrane fraction. These findings suggest the involvement of PKC in the activation of bone marrow-derived macrophages with cisplatin.     

Prognostic indicators for toxic mastitis in dairy cows

OBJECTIVE: To measure serum thyrotropin (TSH), and free thyroxine concentrations in newborns delivered in areas with differing degrees of iodine deficiency, (prior to the salt iodization programme), and to use these biochemical indices to assess the current status of thyroid function in the group. DESIGN: Cross sectional. SETTING: Chemical Pathology, University of Zimbabwe, Medical School. Radio-immunoassay laboratory, Parirenyatwa Hospital. SUBJECTS: 500 healthy full term newborns, aged two to four days old, weighing not less than 2.5 kg. MAIN OUTCOME MEASURES: Thyroid hormone status of newborns. RESULTS: The mean serum FT4 level was found to be slightly but significantly different between the newborns from Harare region, (low goitre prevalent area), and Wedza district (moderate to severe goitre prevalent area). Although there was a trend in the distribution of TSH to higher values in the moderate to severe goitre prevalent areas, the differences were statistically non significant, p = 0.175. The median TSH value in the newborns was 4 microU/ml. TSH values were below 10 microU/ml in 90%, between 10.1 and 20 microU/ml in 9%, and between 20.1 and 32 microU/ml in 1% of the cases. No sex related differences were observed in either the TSH nor the FT4 values of the newborns. CONCLUSION: This study demonstrates that the iodine supplementation programme has been successful, but further monitoring is necessary to ensure complete supplementation throughout the country and to guard against hyperthyroidism which is known to occur during iodine supplementation programmes.     

Non-insulin- and insulin-mediated glucose uptake in dairy cows

Four mid-lactation Holstein dairy cows (mean milk yield on day of experiments 26.1 kg/d) were used in a series of experiments to establish the contribution of non-insulin-mediated glucose uptake to total glucose uptake at basal insulin concentrations. A secondary objective was to determine whether somatostatin affects the action of infused insulin. In part I of the experiment a primed continuous infusion [6,6-2H]glucose (45.2 micrograms/kg per min) was begun at time 0 and continued for 5 h. After 3 h of [6,6-2H]glucose infusion (basal period) a primed continuous infusion of insulin (0.001 i.u./kg per min) was administered for 2 h. Coincidental with the insulin infusion, normal glucose was also infused in order to maintain the plasma glucose concentration at euglycaemia. Part II of the experiment was the same as part I except that somatostatin was infused for 2 h (0.333 micrograms/kg per min) instead of insulin. In part III of the experiment both insulin and somatostatin were infused for the final 2 h. Plasma insulin levels were increased by insulin infusion (to 0.1476 to 0.1290 i.u./l for parts I and III respectively) and were reduced by somatostatin infusion in part II (to 0.006 i.u./l) relative to the basal periods (mean 0.021 i.u./l). Glucose uptake during somatostatin infusion (2.50 mg/kg per min; part II) was 92.0% of that observed in the respective basal period (2.72 mg/kg per min). Circulating insulin levels were much lower than the dose of insulin that causes a half maximal effect on glucose uptake (0.06-0.10 i.u./l for ruminants); consequently insulin-mediated glucose uptake was probably absent in part II. Secondly, glucose uptake following insulin only infusion (4.05 mg/kg per min) was significantly lower than that observed when insulin plus somatostatin was infused (4.69 mg/kg per min), indicating that somatostatin either directly or indirectly enhanced the action of insulin on glucose uptake.