The use of monoclonal antibodies in an enzyme immunospot assay to detect isotype-specific antibody-secreting cells in pigs and chickens

Monoclonal antibodies directed against porcine immunoglobulin isotypes G, G1, G2, M, and A and against chicken immunoglobulin isotopes G, M, and A were tested in an antigen-specific spot-forming cell (SFC) assay based on the principle of the enzyme immunoassay. The SFC assay was used to quantitate ovalbumin (OA)-specific antibody-secreting cells (ASC) in pigs that had been primed and boosted with OA. The SFC assay was also used to quantitate trinitrophenyl (TNP)-specific ASC in chickens that had been primed with TNP-conjugated keyhole lympet haemocyanin (TNP-KLH). Although, the classical plaque-forming cell (PFC) assay cannot reliably detect isotope-specific ASC in pigs and chickens, it can detect these cells in mice. Therefore, we compared the OA- and TNP-specific SFC assays with PFC assays that were specific for these antigens in mice. The study demonstrated that the SFC assay is superior to the PFC assay in detecting both OA-specific ASC and TNP-specific ASC. The frequencies of OA-specific and TNP-specific SFC detected in mice were of the same order of magnitude as those detected in pigs and chickens. We concluded that the SFC assay is the better method for quantitating ASC in pigs, chickens, and probably all domestic animals for which isotype-specific monoclonal antibodies are available.     

Identification of carcass and meat quality quantitative trait loci in a Landrace pig population selected for growth and leanness

The identification of QTL related to production traits that are relevant for the pig industry has been mostly performed by using divergent crosses. The main objective of the current study was to investigate whether these growth, fatness, and meat quality QTL, previously described in diverse experimental populations, were segregating in a Landrace commercial population selected for litter size, backfat thickness, and growth performance. We have found QTL for carcass weight (posterior P > 0.75), cutlet weight (posterior P > 0.99), weight of ham (posterior P > 0.75), shoulders weight (posterior probability > 0.99), and shear firm-ness (posterior P > 0.99) on pig Chromosome 2. Moreover, QTL with posterior P > 0.75 for fat thickness between the 3rd and 4th ribs (Chromosome 7), rib weights (Chromosome 8), backfat thickness (Chromosomes 8, 9, and 10), and b Minolta color component (Chromosome 7) were identified. These results indicate that commercial purebred populations retain a significant amount of genetic variation, even for traits that have been selected for many generations.     

Immunoglobulin G and E responses in various stages of canine leishmaniosis

Pathogenesis in visceral leishmaniosis is associated with depressed cellular immunity and a significant rise of antileishmanial antibodies. We assessed the relative levels of immunoglobulin E anti-Leishmania infantum, together with those of IgG, IgG1 and IgG2, using the enzyme-linked immunosorbent assay (ELISA) test, in non-infected and infected dogs with or without symptoms, and their association with symptoms to differentiate the stages of the infection. The expression of all immunoglobulins (IgG, IgG1, IgG2 and IgE) was higher in symptomatic dogs than in all other categories. IgG and IgG2 expression was higher in the infected asymptomatic group than in the non-infected group, whereas IgG1 and IgE expression was only higher in symptomatic animals. This correlation between the expression of IgG1 and IgE and the pathology of leishmaniosis points to their potential role as markers of the active disease.     

A study on Theileria parva bovis carrier state

In two trials, Theileria parva bovis (which causes ‘January disease’ of cattle in Zimbabwe) produced a carrier state, over the 7–12 months after infection. Very severe clinical reactions were caused by infections from small numbers (29–43) of adult Rhipicephalus appendiculatus ticks, which had engorged on immunized cattle in the field. The transmission from healthy recovered cattle housed indoors was less efficient, even with high numbers of ticks (300). Two out of seven attempts were successful and disease reactions were rather severe. A non-pathogenic Theileria assumed to be Theileria taurotragi was transmitted in three out of seven attempts.     

Cyclooxygenase-2 expression is related with localization, proliferation, and overall survival in canine melanocytic neoplasms

A direct relationship has been firmly established between cyclooxygenase-2 (COX-2) expression and malignant behavior in human melanoma. This report examines the relationship between COX-2 expression and tumor location, mitotic and proliferative indices, degree of T CD3(+) lymphocyte infiltration, overall survival, and frequency of recurrence and metastasis of 57 melanocytic tumors (25 oral and 32 cutaneous). COX-2 was highly or moderately expressed in 88% of oral neoplasms (22 of 25), whereas for their cutaneous counterparts, COX-2 expression was low or insignificant in 75% of cases (24 of 32). High and moderate COX-2 expression levels were observed in 73% of melanocytic tumors with a mitotic index ≥ 3 per 10 high-power fields (26 of 36), whereas in 81% of tumors with a mitotic index < 3 (17 of 21), expression was mild or absent. There were 41 cases with known clinical outcomes; of those showing high, moderate, and mild COX-2 expression, 83.3% (10 of 12), 37.5% (3 of 8), and 25% (2 of 8) died, respectively, whereas 100% of animals showing no COX-2 expression (13 of 13) were still alive at the last follow-up. COX-2 expression was statistically correlated with tumor location, mitotic and percentage Ki-67 proliferative indices, and overall survival, frequency of neoplastic recurrence and metastasis. Regression analysis also showed disease-specific predictive value for COX-2 expression for subjects with melanocytic neoplasms. Additionally, only high COX-2 expression showed significant differences in overall survival, in comparison with moderate, mild, or absent expression. These results suggest that high COX-2 expression may be considered a prognostic biomarker and potentially as a target for therapeutic and preventive strategies in canine melanocytic neoplasms.     

Acaricidal properties of the essential oil and precocene II obtained from Calea serrata (Asteraceae) on the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

Calea serrata Less. (Asteraceae), an endemic species of south Brazil known as "quebra-tudo", is used in Afro-Brazilian religious rituals and in folk medicine for treating liver disorders. Phytochemical studies of the n-hexane extract of this plant demonstrated the presence of precocene II, a benzopyran derivative known for its insecticidal activity. The aim of this work was to isolate this benzopyran and determine the chemical composition of the essential oil of C. serrata and further to evaluate the acaricidal activity of the essential oil and precocene II against the larvae of Rhipicephalus (Boophilus) microplus. The LC(99.9) and LC(50) values obtained with the oil, which presents precocene II and sesquiterpenes, were 3.94 μL/mL and 0.28 μL/mL, respectively. For precocene II this values were 4.25mg/mL and 1.78 mg/mL, respectively. The results indicate a synergistic interaction between the components of the oil and precocene II.     

Culture system and long-term storage of culture media in the in vitro production of bovine embryos

The optimum culture system for in vitro matured and fertilised oocytes still remains to be clarified. Culture media (CM) for mammalian embryos are routinely prepared fresh for use and preserved under refrigeration during one or two weeks. The purposes of this work were (1) to compare the efficiency of a synthetic oviduct fluid (SOF) with two different bovine serum albumin (BSA) concentrations (3 and 8 g/L) for the in vitro production of bovine blastocysts, (2) to test the effect of timing on adding fetal calf serum (FCS) to the SOF, and (3) to evaluate the effects on bovine embryo development of freezing and lyophilisation as procedures for preserving the SOF. Supplementation of SOF with 3 g/L BSA increased Day-7 blastocyst expansion rates (18.3 ± 1.6 vs. 14.4 ± 0.7; P < 0.05), although no differences in hatching rates were found. Addition of FCS to SOFaa (SOF with amino acids) medium supplemented with sodium citrate (SOFaaci) at 48 and at 72 h post-insemination (PI) allowed obtaining higher Day-6 embryo development rates than when FCS was added at 18 or 96 h PI (Day-6 morulae + blastocyst rate: 30.0 ± 1.1, 40.8 ± 1.1, 43.9 ± 2.3 and 39.3 ± 0.5 for FCS addition at 18, 48, 72 and 96 h, respectively). Hatching rates were significantly improved when serum was added at 72 h PI. Finally, both refrigeration and lyophilisation appeared as useful cryopreservation procedures for SOFaaci, although a significant loss of its ability to support embryo development, compared to the control fresh culture medium, was observed.     

Metabolic changes induced by regular submaximal aerobic exercise in meat-type rabbits

Pannon White growing rabbits (a group of 8) were exposed to treadmill exercise (3-9 m/s, 1.2-1.6 km/day) twice a day for 4 weeks, while additional 8 animals, kept inactive, were assigned as the control group. Weekly, 12 hours after exercise, venous blood was taken for serum metabolite and enzyme activity measurements. Total serum protein, albumin and creatinine levels significantly increased during the second half of the training, as compared to the control group. Triacylglycerol levels in the exercised group as compared to controls, however, were higher only after the first and the fourth weeks of the experiment. Resting non-esterified fatty acid (NEFA) concentration of the trained rabbits was lower at the end of the trial. On the other hand, there were no significant differences, as compared to the respective controls, in serum urea, total and HDL cholesterol levels. At the end of the exercise alkaline phosphatase activity was higher and total lactate dehydrogenase activity was lower in the trained rabbits. Serum alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl transpeptidase activities were not changed, while creatine kinase activity was slightly lower in the trained group. The serum cortisol concentration was not different in the trained and control rabbits.     

Pregnancy Rate in Zebu Cows with Two Different Postpartum Intervals Exposed to a Two-Bull Rotational System

In order to evaluate the reproductive effect of a rotational breeding system, the pregnancy rate of 69 Zebu cows was assessed, 32 late postpartum (211±93 days postpartum, LP) and 37 early postpartum (averaging 63±8 days after calving, EP). In the LP group, 60% (19/32) were cycling before exposure to the bulls but only 38% (14/37) in the EP group. The two groups were dissimilar from the start (p<0.05). The cows were exposed in a rotational breeding system, with six Brahman bulls with previous sexual experience paired off in three groups (A–B, C–D and E–F). The cows were exposed to each pair of bulls for a period of 3 weeks, with one week in between each period, when the females were without the presence of a male. In the LP group, the pair of bulls A–B obtained a pregnancy rate of 72% (23/32), whereas for bulls C–D the pregnancy rate was 33% (3/9) and for bulls E–F the pregnancy rate was 67% (4/6). In contrast, for the EP cows, bulls A–B obtained a pregnancy rate of 32% (12/37), bulls C–D a pregnancy rate of 67% (12/18) and for bulls E–F a pregnancy rate of 55% (6/11). The percentage pregnancy obtained using the pair of bulls A–B differed (p<0.05) between LP and EP, whereas for bulls C–D and E–F it was similar. The pregnancy rate was different (p<0.05) in the first 3 weeks of mating for the LP group, the highest number of cows becoming pregnant during the first 3 weeks of the study. In contrast, the highest number of pregnancies in EP occurred evenly during the last 6 weeks. This study suggests that the reproductive performance of pairs of bulls in a 9-week rotational programme with the overall pregnancy rate is similar (94% in the LP and 81% in the EP). However, the time taken for the females to become pregnant, and hence the performance of the bulls, is related to the average number of days postpartum for the cows.     

Comparative fluorescein angiography of the normal sheep and goat ocular fundi

Fluorescein angiography without sedative or anesthetic agents was evaluated in 20 normal goats and 20 normal sheep. All of the angiographic phases were observed using 20 mg/kg fluorescein IV in both species. Fundus fluorescein angiography results revealed wide stars of Winslow in the tapetal fundus, central or marginal flow during the first part of the arterial phase, delayed filling of the focal areas in the choroid near the optic disc that often coincided with others in the disc, and lack of evidence of the 'striate area' in the tapetal fundi. In goats, the angiographic times were 6.54+/-1.25 s for the arterial phase (TA), 7.80+/-1.37 s for the arterio-venous phase (TAV), and 14.13+/-2.01 s for the venous phase (TV). I1: 1.30+/-0.30 s (time elapsing between TA and TAV), and I2: 6.20+/-1.60 s (time elapsing between TAV and TV). In sheep, times were 9.54+/-2.18 s TA, 11.73+/-2.10 s TAV, and 20.86+/-2.74 s TV. I1: 2.04+/-0.75 s and I2: 8.98+/-2.47 s, respectively. Due to the large size of the fundic vessels in sheep and goats, fluorescein angiography of the retinal vasculature can facilitate the study of the different vascular diseases in these species.