Production and preliminary characterization of monoclonal antibodies to chicken anemia agent

Mice were immunized with partially purified preparations of the Cux-1 isolate of chicken anemia agent (CAA), and their splenocytes were fused with NSO myeloma cells. Three patterns of staining of CAA-infected cells were recognized when the resulting hybridomas were screened by indirect immunofluorescence (IIF). Hybridomas representative of each staining pattern were cloned, and the monoclonal antibodies (MAbs) were characterized. Type 1 staining was indistinguishable from that produced by polyclonal chicken antisera to CAA. Type 2 staining was confined to large nuclear inclusions. Type 3 staining was predominantly nuclear and granular, and differed from type 1 in being more intense and occurring in a higher proportion of nuclei. Three MAbs producing type 1 staining were predominantly Cux-1-specific by IIF; they also reacted to lower titers with the Gifu-1 isolate but not at all with three other CAA isolates. These MAbs had very slight neutralizing activity against Cux-1. Another MAb giving type 1 staining reacted with all CAA isolates tested to high titers in IIF and neutralization tests. MAbs with type 2 and type 3 staining reacted by IIF with all CAA isolates tested but possessed no neutralizing activity. The availability of MABs to CAA should facilitate development of diagnostic tests for the virus.     

Modelling the impacts of agriculture in mixed-use landscapes: a review and case study involving two species of dabbling ducks

Context

This study synthesizes evidence from three separate surveys of American Black Duck and Mallard breeding habitat usage to quantify the effects of agriculture at the landscape scale.

Objectives

To assess duck breeding activity in agricultural landscapes within the Canadian maritimes in order to measure the overall impact of agricultural land use, the response to particular agricultural activities, and the influence of landscape configuration.

Methods

Models, constructed using a long-term census (SURVEY1), were used to predict habitat selection for two other independent surveys (SURVEY2, SURVEY3). Predictions incorporated information about wetland area and diversity, as well as anthropogenic factors, allowing subsequent analyses to focus on the remaining residual error attributable to agricultural effects.

Results

SURVEY2 results demonstrated that the proportion of active agriculture is an important indicator of the severity of human disturbance, yielding threshold estimates of 39% for Mallards and 60% for Black Ducks, with an overall average of 50%. Agricultural conversion beyond these thresholds deterred breeding ducks independently of other factors. SURVEY3 land cover information demonstrated that the presence of cropland intensified this deterrence effect, even at levels as low as 10%. Woodland cover (in excess of 30%) was important for both species, but its configuration was not.

Conclusions

In addition to quantifying threshold effects, this study reaffirms that woodland is an important part of the maritime landscape matrix, and contributes positively to habitat diversity in mixed use, moderate intensity agricultural regions. Wetland restoration in agricultural landscapes should monitor and promote less crop-intensive, mixed-use practices.

     

Lectin histochemical pattern on the normal and cystic ovaries of sows

The lectin histochemical pattern (LHP) was characterized and compared in normal and cystic ovaries of sows. Six biotinylated lectins (PNA, SBA, WGA, RCA‐1, DBA and UEA‐1) were used on tissue sections. In the normal ovaries, the reaction to UEA‐1 and SBA was mild to moderate in mesothelial and endothelial cells. RCA‐1 staining was mild to moderate in theca interna of growing follicles, corpora luteum and mesothelium. In addition, this lectin presented strong reaction in endothelial cells, granulosa cells of atretic follicles, zona pellucida of growing follicles and plasma. DBA showed strong intensity in mesothelial and endothelial cells. There was mild to moderate reactivity to WGA in granulosa cells, corpus luteum and theca interna of follicles in development, and moderate in zona pellucida, in granulosa cells of atretic follicles and mesothelium. PNA staining was mild to moderate in oocytes and in the adventitia and media of medullary arteries. Changes in the LHP of the cystic ovaries were noted; however, there were no differences in these findings between the follicular and luteinized cysts. UEA‐1 reactivity in the cystic ovaries was moderately reduced in the mesothelial and endothelial cells, whereas there was mild reduction in the DBA staining in the granulosa cells. Reaction to RCA‐1 and WGA in the cysts also was decreased in theca interna, zona pellucida and granulosa cells of atretic follicles. Furthermore, endothelium and theca interna in the cystic ovaries presented mild reduction of marcation to SBA, whereas there was decreased reactivity to PNA in the oocytes and adventitia and media layers of the medullary arteries. The results of the current study show that cysts modify the LHP in swine ovaries. These changes of glycoconjugates in many ovarian structures could modify diverse process and may be one of the reasons for decreased fertility in sows.     

Molecular genotyping of multinational ovine and caprine Corynebacterium pseudotuberculosis isolates using pulsed-field gel electrophoresis

Caseous lymphadenitis (CLA) is a chronic, suppurative disease, with a worldwide distribution, caused by Corynebacterium pseudotuberculosis. The clinical manifestation of CLA is known to vary between different countries, and has been postulated to be due to differences in the strains present in these countries. Forty-two sheep and goat isolates of C. pseudotuberculosis from Australia, Canada, Eire, The Netherlands and Northern Ireland were characterized by pulsed-field gel electrophoresis (PFGE), biotyping, antimicrobial susceptibility, and production of phospholipase D. The PFGE-determined genotypes of this multicentric collection were then compared with representative ovine and caprine isolates from a previously published panel of PFGE profiles of United Kingdom isolates. Digestion with SfiI generated 16-18 bands in the 48.5 and 290 kb range, and differentiated four distinct pulsotypes amongst the 36 ovine and 6 caprine strains which displayed remarkable homogeneity. Based on these results, it would appear that the genome of C. pseudotuberculosis is highly conserved, irrespective of the country of strain origin.     

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.     

Bovine trypanosomiasis in Southern Tanzania: Parasitological and serological survey of prevalence

In a survey for bovine trypanosomiasis blood smears from 1,617 cattle at 72 sites were examined. Trypanosomes were found in 93 cattle, representing 16% of the cattle in herds in which trypanosomiasis was confirmed. Of the positive cattle 56% had infections with T. congolense, 17% T. vivax and 2.2% T. brucei. Five cattle had mixed infections and in 18 cattle the species was not identified. Sera from 1,352 cattle were tested using microelisa. Ten out of 16 sites, at which no trypanosomes were found in blood smears and at which trypanocides were in use, had over 15% seropositive cattle compared with five of 19 sites at which trypanocides were not in use. It was concluded that the microelisa was a useful aid to the diagnosis of bovine trypanosomiasis and that there is a need for accurate records of drug use and livestock movements to be kept. The serious risk of drug resistant strains of trypanosomes emerging due to the uncontrolled use of trypanocides is emphasised.     

Pre-breeding Ovaro–Uterine Ultrasonography and its Relationship with First Service Pregnancy Rate in Seasonal-Calving Dairy Herds

The objectives of the study were to characterize an ultrasound reproductive tract scoring (URTS) system to assess suitability for breeding in dairy cows, to describe the prevalence of these scores in commercial dairy herds and to examine their relationship with subsequent fertility. Ultrasound examinations (7797) were performed on 5751 Holstein–Friesian cows prior to breeding in 62 seasonally calving herds over 2 years. Data recorded from images of both ovaries and the uterus were combined into a six point scoring system and the prevalence of cows with cystic ovarian follicles and uterine abscesses and adhesions was recorded separately. The prevalence of ovulatory cows which had completed uterine involution (score 1), or had mild (2) or moderate endometritis (3) was 62.5%, 21.7% and 2.2%, respectively. The prevalence of anovulatory cows with moderate endometritis (4), ovulatory cows with pyometra (5) and anovulatory cows which had completed uterine involution (6) was 3.3%, 2.2% and 8.1%, respectively. The interval between calving and examination differed between cows with each of the scores 1, 2, 5 and 6 (61, 46, 53 and 50 days, respectively, p < 0.05) but not between cows with scores 3 and 4 (37 and 35 days, respectively). Ovulatory cows which had completed uterine involution (score 1) had a higher likelihood of pregnancy to first service than ovulatory or anovulatory cows which had not completed uterine involution (p ≤ 0.10). The prevalence of cows with cystic ovarian follicles or abscesses or adhesions of the reproductive tract was 3.9% and 1.2%, respectively. In conclusion, 29% and 11% of cows in seasonally calving and breeding dairy herds had not completed uterine involution or were anovulatory prior to the mating start date, respectively. Both conditions, detected using a URTS system, significantly reduced first service pregnancy rate in these pasture-based dairy herds.