Effect of thermal exposure on physiological adaptability and seminal attributes of rams under semi-arid environment

Thermal stress in hot semi-arid environment is a major limitation of sheep production in tropical and sub-tropical climatic condition. The animals tend to maintain homeostasis through physiological adjustments in a hot environment (maximum temperature reaches up to 47.5 °C). Therefore, the present study was carried out to assess the effect of thermal exposure on physiological adaptability and seminal attributes of rams under semi-arid environment. The experiment was conducted for eight weeks involving sixteen Malpura crossbred rams (GMM: Garole X Malpura X Malpura). The rams were divided equally into two groups, designated as G1 and G2, respectively. The rams in G1 (Control) group were kept in a sheep shed under naturally prevailing environment without artificial manipulation of ambient temperature (Temperature 30.48±0.38 °C; Relative Humidity 28.59±1.15%). The rams of G2 group were exposed to different temperature at different hours of the day (38 °C at 1000–1100 h; 40 °C at 1100–1200 h; 42 °C at 12:00–1300 h; 43 °C at 1300–1400 h; 44 °C at 1400–1500 h and 42 °C at 1500–1600 h) in a climatic chamber for thermal exposure. Physiological responses, blood biochemical profile, blood endocrine profile, sexual behavior and seminal attributes were measured for both the groups. Thermal exposure significantly (P<0.05) increased the water intake; respiration rate, rectal temperature and skin temperature at afternoon in rams. Exposure of rams to thermal stress (G2) significantly (P<0.05) increased cortisol level and decreased tri-ido-thyronine level. The latency period after the first ejaculation, decreased significantly (P<0.05) in G2. The percentage of rapid motile sperm, linearity and average path velocity of sperm were also altered significantly (P<0.05) in thermal exposed rams as compared to control. However, comparable feed intake, body weight, and major blood biochemical parameters, as well as acceptable semen quality attributes of all the rams indicated that the Fec B gene introgressed Malpura cross rams adapted to the thermal exposure under semi-arid tropical climate.     

Antibody-based diagnosis of small ruminant lentivirus infection in seminal fluid

Antibody-based diagnosis of small ruminant lentiviruses (SRLVs) has been efficiently achieved using serum and milk, but not semen, for which polymerase chain reaction (PCR) has been proposed as a confirmatory technique. This work, involving 296 ovine (Ovis aries) and caprine (Capra hircus) semen donors, investigates whether seminal fluid (SF) can be reliably used in antibody-based SRLV diagnosis. First, a gold standard was established to assess the infection status and determine the sensitivity and specificity of three commercial enzyme-linked immunosorbent assays (ELISAs) in serum testing using Western blot and PCR as confirmatory tests. For SF testing, both gold standard and serum testing results were used as reference. The performance of SF testing was affected not only by the ELISA assay sensitivity (related to antigen spectrum) compared with that of the gold standard (as it occurred in serum testing) but also by SF sample quality and SF working dilution. Nonturbid SF samples, commonly collected in artificial insemination centers (AICs), were required. Compared with serum, SF testing had a decreased sensitivity in two of the ELISA assays (with original serum working dilutions ≤1/20 in serum testing) but reached a similar sensitivity (and specificity) in the assay designed to work at the highest serum dilution (1/500). A SF concentration of about 1/2 (250-fold that used in serum testing) was found optimal in this assay, yielding highly repeatable results that were in almost perfect agreement with those of serum testing (κ ± SE, 0.91 ± 0.81). Thus, SF ELISA can be reliably applied in antibody-based SRLV diagnosis. This information may be useful to control infection in AICs and animal and semen trade programs requiring health-certified quality of semen donors.     

Peptidase activities in the semen from the ductus deferens and uterus of the neotropical rattlesnake Crotalus durissus terrificus

To understand the role of peptidases in seminal physiology of Crotalus durissus terrificus, intra- and inter-seasonal activity levels of acid (APA), basic (APB), puromycin-sensitive (APN-PS) and puromycin-insensitive neutral (APN-PI), cystyl (CAP), dipeptidyl-IV (DPPIV), type-1 pyroglutamyl (PAP-I) and prolyl-imino (PIP) aminopeptidases as well as prolyl endopeptidase (POP) were evaluated in soluble (SF) and/or membrane-bound (MF) fractions of semen collected from the ductus deferens of the male reproductive tract and from the posterior portion of the uterus. Seminal APB, PIP and POP were detected in SF, while other peptidases were detected in SF and MF. Only the convoluted posterior uterus in winter and autumn had semen. Relative to other examined peptidases, in general, APN-PI, APN-PS and APB activities were predominant in the semen from the uterus and throughout the year in the semen from the ductus deferens, suggesting their great relevance in the seminal physiology of C. d. terrificus. The levels of peptidase activities in the ductus deferens semen varied seasonally and were different from those of semen in the uterus, suggesting that their modulatory actions on susceptible peptides are integrated to the male reproductive cycle events and spermatozoa viability of this snake.     

Evaluation of the presence of equine viral herpesvirus 1 (EHV-1) and equine viral herpesvirus 4 (EHV-4) DNA in stallion semen using polymerase chain reaction (PCR)

In the horse, the risk of excretion of two major equine pathogens (equine herpesvirus types 1 (EHV-1) and 4 (EHV-4)) in semen is unknown. The objective of our study was to assess the possible risks for the horizontal transmission of equine rhinopneumonitis herpesviruses via the semen and the effect of the viruses on stallion fertility.Samples of stallion semen (n = 390) were gathered from several different sources. Examination of the semen involved the detection of viral DNA using specific PCR. The mean fertility of the stallions whose sperm tested positive for viral DNA and the mean fertility of stallions whose sperm did not contain viral DNA, were compared using the Student's t-test.EHV-4 viral DNA was not detected in any of the semen samples. EHV-1 DNA was identified in 51 of the 390 samples, (13%). One hundred and eighty-two samples came from 6 studs and there was significant difference (p < 0.05) among the proportion of stallions whose semen tested positive for viral DNA from 0 to 55% between the studs.There was a significant difference (p < 0.014) between the fertility of stallions whose semen tested positive for viral DNA and those whose semen was free from viral DNA. The stallions that excreted the EHV-1 virus in their semen appeared to be more fertile than the non-excretors, but this difference was in fact related to the breeding technique since higher proportion of excretors were found among those whose semen was used fresh rather than preserved by cooling or freezing.In conclusion, this study suggests that the EHV-1 virus may be transmitted via the semen at mating or by artificial insemination as demonstrated with other herpes viruses in other species.     

Flow cytometric evaluation of sperm apoptosis in semen of silver foxes in the breeding period

The objective of the study was to evaluate cytometrically the percentage of apoptotic and necrotic spermatozoa in fresh semen of silver foxes in the breeding season. In males F3 and F4 with high percentages of early apoptotic (A+Pi−), late apoptotic (A+Pi+) and necrotic (A−Pi+) spermatozoa as well as 56–65% of living spermatozoa (A−Pi−) with progressive motility, the semen was characterised by reduced fertility. In males F1 and F2 with spermatozoa showing the motility and viability of 89–90% and high percentages of living cells that do not bind Annexin V and propidium iodide, the semen was assessed as valuable and useful for artificial insemination. Amongst 16 females of group I and II inseminated with semen from F1 and F2 males, 15 (93.75%) had multi-cub litters – on average 6.1 and 4.8, respectively. In contrast, amongst 16 females of group III and IV inseminated with semen from F3 and F4 males, only 10 (62.5%) had litters with few cubs (on average 2.6 in group III and 2.1 in group IV). Our findings explicitly indicate that semen of farm male foxes should be evaluated before the breeding season, as one of the causes of reproduction failures is likely to be a high percentage of apoptotic and necrotic spermatozoa. Thanks to flow cytometry, fresh ejaculates can be speedily evaluated and their usefulness for artificial insemination determined.     

Distribution of heavy metals in surface sediments of the Bacochibampo Bay,Sonora, Mexico

Abstract

The aim of this study was to determine the levels of heavy metals (Al, Cd, Cr, Cu, Fe, Mn and Pb) in the geochemical fractions of the coastal surface sediments from the Bacochibampo Bay in Sonora, Mexico. Two surveys were conducted (March and September) during 2004, at eight sampling stations inside the bay, and in three natural effluents discharged into this bay. The extraction of metals was carried out using a microwave oven method and the quantification was done by atomic absorption spectro-photometry. The highest detected concentration of total heavy metals in sediments was: Fe>Al>Mn>Pb>Cr>Cu>Cd, with the following concentration values: Fe (1.72%), Al (1.03%), Mn (416.31 mg kg^?1), Pb (11.73 mg kg^?1), Cr (11.41 mg kg^–1), Cu (6.78 mg kg^–1) and Cd (1.33 mg kg^–1). The levels of total heavy metals (Al, Cr, Cu, Fe, Mn and Pb) were much less than the lowest observable effect level (LEL) which indicates that the sediments were not from polluted areas and that the origin of the metals was due to natural conditions. However, concentrations of Cd were much higher than the low effect level (LEL), over 40% of metal was detected in the exchangeable fraction and carbonates. The normalisation study showed a high degree of enrichment of Cd in all the sampling stations in the Bacochibampo Bay (samples EF 34–87) and in the natural flows that discharge into this bay (samples EF 22–35%), which exceeds by several orders of magnitude the value of sample EF 1, which indicates that Cd is anthropogenically induced. Based on these results, it is important that precautionary measures are established, since the deposited Cd in these fractions may be potentially toxic, due to the physicochemical changes that occur in the environment. Thus, future studies will focus on identifying problems involved with Cd bioaccumulation in different trophic levels.     

The availability of soil boron fractions to olive trees and barley and their relationships to soil properties

Soil boron occurring in various forms was correlated with boron contents in the leaves of olive trees in 51 and in barley leaves in 20 soils. The amounts of boron: in soil solution (CwsB), non specifically adsorbed (NsaB), specifically adsorbed (SaB), occluded in Mn oxides (MnoB), occluded in amorphous Fe-Al oxyhydroxides (FeoB), were correlated with soil properties such as: organic matter content, pH, free aluminum and iron oxyhydroxides (Ald, Fed), amorphous aluminum and iron oxyhydroxides (Alo, Feo). The later correlation studies were conducted on a total number of 153 soil samples inclusive of the soils used for the plant uptake investigations.The results show that the boron contents in the leaves of olive trees were significantly correlated with FeoB, CwsB, SaB and MnoB but with CwsB, NsaB, SaB and FeoB in barley leaves. The respective correlation coefficients suggest that available forms of soil boron vary with plant species.The hot water soluble boron was significantly correlated with all the fractions of boron studied except MnoB, confirming its value as a measure of available soil boron. However, the non significant (a=0.05) correlation with MnoB, which show significant correlation with the contents in the leaves of olive trees and barley, suggest that this procedure does not extract significant amounts of available B held in Mn oxyhydroxides.The highest amount of CwsB originated from the Cws B and to a lesser degree from the SaB and NsaB. The correlation among the boron held in the forms studied and the selected soil properties were either no significant or significant with the highest correlation found between FeoB and pH and Feo.     

Soil phosphorus fractions,aluminum, and water retention as affected by microbial activity in an Ultisol

Increased organic matter input into weathered and infertile soils through agricultural techniques such as minimum tillage or agroforestry can improve P availability to crops. Organic matter is an energy source for microbes, and their activity may be responsible in part for increased levels of labile P. The objective of the work reported here was to examine, in a highly weathered Ultisol, the influence of microbial activity in mobilizing P, maintaining it in a plant-available state, and preventing its fixation, and the effect of N and biocides on these processes. Exchangeable aluminum and soil moisture were also determined, since they interact with microbes and soil P.Results showed that increased microbial activity reduced sorption of dissolved and organic P by soil, maintained inorganic P in soluble and labile pools, increased microbial P, decreased mineral P, increased exchangeable Al, and increased water retention. Additions of N and biocides had variable effects, probably due to complex interactions between N, degrading biocides, and microbial populations.     

The response of Glomus fistulosum–maize mycorrhiza to treatments with culture fractions from Pseudomonas putida

 This study examined which culture fraction of the plant-growth-promoting bacterium Pseudomonas putida (Trevisan) Migula has an effect on growth and mycorrhiza formation of maize (Zea mays L.). Shoot dry weight and total leaf area of plants did not increase after inoculation with Glomus fistulosum but were significantly higher than the controls when the plants were dualinoculated with G. fistulosum and living cells of P. putida. Mycorrhizal infection of the roots was significantly higher when plants were inoculated with G. fistulosum together with living cells of P. putida or with G. fistulosum and dialysed cell extracts of P. putida than with G. fistulosum alone. Development of arbuscular mycorrhizal (AM) extraradical hyphae and the proportion of extraradical hyphae showing NADH diaphorase activity were significantly enhanced by inoculation of plants with living cells of P. putida or dialysed cell extracts of P. putida. No stimulation of extraradical hyphae proliferation from in vitro incubated mycorrhizal root segments was observed after application of culture fractions of P. putida. However, the percentage contamination of the root segments by extraneous filamentous fungi significantly decreased in the presence of livingcells of P. putida. Accepted: 12 January 1996