Three cases of disputed paternity in dogs resolved by the use of DNA fingerprinting

DNA fingerprinting with the probes 33.15 and alpha-globin 3'HVR has been used to resolve three cases of disputed paternity in dogs. For each pedigree it was necessary to establish which bands in the DNA fingerprints of the offspring were of paternal origin, and then establish which putative sire carried all these bands. In the first case, a litter of Rhodesian Ridgebacks, twelve DNA bands were informative in establishing paternity. In the second case, a litter of Afghan hounds, five DNA bands established paternity, Lastly, in a litter of Border collies, five DNA bands established paternity. In each case a single dog only sired the entire litter.     

In situ provision of drinking water to grazing dairy cows improves milk production

AIMS: To determine the effect of providing water within the area grazed by dairy cows on milk yield and quality, compared to requiring cows to walk to a distant water trough, on a dairy farm in the Pampa region of Argentina during summer.

METHODS: Holstein dairy cows were allocated to two herds with similar parity, days in milk and milk production. They were grazed in one paddock that was divided in two, with a fixed water trough at one end. Cows were moved twice daily to grazing plots within the paddock. Control cows (n=66) could only access water from the fixed trough, whereas supplemented cows (n=67) also received water from a mobile trough within the grazing plot. Milk production of each cow, and water consumption of the two herds were measured daily over 62 days. Milk composition for each herd was determined weekly from Days 18 to 60 of the study, and grazing behaviour was observed between 08:00 and 16:00 hours on Days 11–15, 19–22 and 39–43.

RESULTS: Over the 62 days of the study, supplemented cows produced 1.39 (SE 0.11) L/cow/day more milk than Control cows (p=0.027). Estimated mean daily water intake was 50.4 (SE 2.1) L/cow/day for supplemented cows and 58.2 (SE 2.7) L/cow/day for Control cows (p=0.004). Percentage total solids in milk was higher for supplemented (12.5 (SE 0.06)%) than Control (12.4 (SE 0.04)%) cows (p=0.047). During the periods of behavioural observation, a higher percentage of cows in the water supplemented than the Control herd were observed in the grazing area (p=0.012).

CONCLUSIONS AND CLINICAL RELEVANCE: This preliminary study demonstrated that provision of water to dairy cows within the grazing plot was beneficial for milk production and composition, and may be associated with longer periods spent within the grazing area, during hot weather in the Pampa region of Argentina.     

Spermatic and oxidative profile of domestic cat (Felis catus) epididymal sperm subjected to different cooling times (24, 48 and 72 hours)

Cooling stored epididymal samples for several days allows facilities to transport and process genetic material post‐mortem. Improvements to this practice allow the preservation of sperm from domestic cats, which are the ideal study model for wild felids. However, the modifications in spermatic features and the oxidative profile are not fully understood in cats. This information is necessary for the development of biotechniques, such as new extenders for cryopreservation. Therefore, the purpose of this study was to evaluate the spermatic and oxidative profile in samples from the epididymal cauda of domestic cats cooled at 5°C for 24, 48 and 72 hr. Spermatozoa were collected from the epididymis cauda. Evaluations consisted of computer‐assisted sperm analysis (CASA), plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, sperm DNA integrity (toluidine blue), mitochondrial activity (3′3 diaminobenzidine), activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD), measurement of lipid peroxidation (TBARS) and protein oxidation. A decrease in sperm motility parameters was observed after 72 hr of cooling (i.e. total and progressive) with a higher percentage of minor (37.7 ± 6.3%) and total defects (53.4 ± 6.3%). Additionally, a decrease in high mitochondrial activity (Class I: 16.6 ± 2.2%) occurred after 72 hr. The decrease in motility rates after a long cooling time probably was caused by the increase in sperm abnormalities. A long cooling time causes cold shock and mitochondrial exhaustion, but there was no observed change with the oxidative stress condition. Therefore, cat epididymal sperm stored at 5°C appear to maintain a high quality for up to 48 hr of cooling time.     

Pregnancy‐Induced ISG‐15 and MX‐1 Gene Expression is Detected in the Liver of Holstein–Friesian Heifers During Late Peri‐Implantation Period

The bovine embryonic signal interferon‐τ (IFN‐τ) produced by the trophoblast is known to pass through the uterine fluid towards the endometrium and further into the maternal blood, where IFN‐τ induces specific expression of interferon‐stimulated gene expression (ISG), for example in peripheral leucocytes. In sheep, it was shown experimentally by administration of IFN‐τ that ISG is also detectable in the liver. The objective was to test whether ISG can be detected in liver biopsy specimens from Holstein–Friesian heifers during early pregnancy. Liver biopsies were taken on day 18 from pregnant and non‐pregnant heifers (n = 19), and the interferon‐stimulated protein 15 kDa (ISG‐15) and myxovirus‐resistance protein‐1 (MX‐1) gene expression was detected. The expression of both MX‐1 (p: 24.33 ± 7.40 vs np: 9.00 ± 4.02) and ISG‐15 (p: 43.73 ± 23.22 vs 7.83 ± 3.63) was higher in pregnant compared to non‐pregnant heifers (p < 0.05). In conclusion, pregnancy induced ISG‐15 and MX‐1 gene expression in the liver already at day 18 in cattle.     

Does returning sites of historic peri‐urban waste disposal to vegetable production pose a risk to human health? – A case study near Manchester,UK

Urban waste disposal occurred on fenland to the west of Manchester, England, between 1900 and 1964. The reclaimed fenland, Chat Moss, is now used for mixed arable farming. A total of 1.92 Mt of waste including privy midden, street sweepings, clinkers and slaughterhouse refuse was incorporated into the moss resulting in a modified topsoil with raised pH and reduced organic matter content compared with the subsoil. Elevated levels of potentially toxic elements (PTEs) are observed in the topsoil beyond the typical depth of atmospheric contamination; Cd and As concentrations exceed soil guideline values (SGVs) at 1.8 and 43 mg/kg, respectively. Sequential extraction indicates that waste‐derived Pb, Zn and Ni remain predominantly in the residual fraction, whereas Cu was mainly organically bound. Arsenic was predominately found in oxide and organic matter fractions with Cd in carbonate, oxide, organic matter and residual fractions. Pot trials indicated limited uptake of PTEs by vegetables grown on the waste‐amended soil, with the exception of Cd uptake by lettuce (0.22 mg/kg FW) and Pb uptake by radish (0.16 mg/kg FW), which exceeded current EU limits of 0.2 and 0.1 mg/kg FW, respectively. Hazard quotients (HQs) identified no risks to adults from consumption of vegetables grown in these soils with the exception of lettuce consumption with a HQ of 1.4. Risks to children were slightly greater with HQs >1 for Cd in lettuce, spinach, carrots and onion, As in lettuce, parsley and onion and for Zn in spinach.     

Systemic accumulation of novel proteins in the apoplast of the leaves of potato plants following root invasion by the cyst-nematode Globodera rostochiensis

Systemic changes in gene expression have been analysed in potato plants following root infection with the cyst nematode Globodera rostochiensis. Root invasion by the pathogen induced major changes to occur in the composition of proteins extracted in intercellular fluid from the leaf apoplastic space. The changes involved the disappearance of innate leaf proteins and the accumulation of novel gene products. Analysis by SDS-PAGE indicated that the leaf polypeptides induced on nematode invasion are not cultivar or nematode pathotype-specific. Aspirin treatment of leaves induced a similar pattern to that induced systemically by nematodes, but several gene products were specific for the latter method of induction. In particular, two abundant [¹²⁵I]-Con A binding polypeptides of high M_r accumulated in intercellular fluid only on aspirin treatment. The results are discussed in the context of the role of the extracellular leaf proteins and their modulation by systemic signals.     

Baculoviruses as vectors for foreign gene expression in insect cells

Recombinant DNA technology has enabled the expression of a large number of eukaryotic genes in vitro. There has been growing interest in recent years in the development of baculovirus expression vectors as an easily manipulated gene expression system. Both virus and insect cell culture are relatively easy to handle, and biologically active proteins have been produced abundantly from a variety of eukaryotic, prokaryotic and viral genes. This review describes the molecular biology of baculoviruses and some of the current applications of the baculovirus expression system in insect cells. Specific emphasis is placed on those features relevant to the use of this system in pesticide research.     

Seroprevalence of Neospora caninum infection following an abortion outbreak in a dairy cattle herd

OBJECTIVE: To investigate the seroprevalence of Neospora caninum infection in a commercial dairy cattle herd, 15 months after detection of an abortion outbreak. PROCEDURE: Sera from the whole herd (n = 266) were examined for N caninum antibodies by indirect fluorescent antibody test (IFAT) and immunoblot analysis. Herd records were reviewed to collate serological results with abortion history, proximity to calving, and pedigree data. RESULTS: The seroprevalence of N caninum infection was 24% (63/266) for IFAT titre > or = 160, 29% (78/266) for immunoblot positive (+ve), and 31% (82/266) for IFAT > or = 160 and/or immunoblot +ve; 94% (59/63) of animals with IFAT > or = 160 were immunoblot +ve. The association between seropositivity (IFAT > or = 160 and/or immunoblot +ve) and history of abortion was highly significant (P < 0.001); the seroprevalence was 86% (18/21) in aborting cows, compared with 30% (50/164) in non-aborting animals. The abortion rate for seropositive cows was 26% (18/68) compared with 3% (3/117) for seronegative animals. IFAT titres of infected cows were higher within 2 months of calving than at other times (P < 0.001). The association between seropositivity in dams and daughters was highly significant (P = 0.009). CONCLUSIONS: The abortions were associated with N caninum infection and there was evidence of reactivation of latent infection close to calving and congenital transmission of infection. Immunodominant antigens identified by immunoblots may prove useful for improved diagnostic tests.     

Noninvasive monitoring of stress hormone levels in a female steller sea lion (Eumetopias jubatus) pup undergoing rehabilitation.

Steller sea lions (Eumetopias jubatus) rarely strand in areas monitored by humans, and there is little published data on the diseases, parasites, nutritional state, and stress levels of Steller sea lions in the wild. In May 2002, a female Steller sea lion pup (EJS-02-01) was sighted separated from her mother after strong storms in Southeast Alaska. After 5 days of observations, EJS-02-01 was transferred to the Alaska SeaLife Center (ASLC) in Seward, Alaska. During 11 mo of rehabilitation at ASLC, body weight was monitored and opportunistic fecal samples (n = 86) were analyzed for corticosterone concentrations. Fecal corticosterone concentrations ranged from 15 to 3,805 ng/ g for EJS-02-01. Peak corticosterone values reflected responses to acute stressors during rehabilitation. EJS-02-01 was successfully released at Gran Point, Alaska, in April 2003. Fecal corticosterone assay monitoring provided a valuable tool to monitor various stressors and is useful in monitoring long-term situations like rehabilitation.