A sandwich enzyme immunoassay for bovine interferon-γ and its use for the detection of tuberculosis in cattle

An in vitro cellular assay for bovine tuberculosis has recently been developed. This assay detects gamma-interferon released in response to specific antigen in a whole blood culture system. The bio-assay previously described for the detection of bovine gamma-interferon (IFN-gamma) has now been replaced with a sandwich enzyme immunoassay (EIA) which utilises two monoclonal antibodies to bovine IFN-gamma. The EIA detects less than 25pg/ml of recombinant bovine IFN-gamma and is specific for biologically active bovine IFN-gamma; and does not detect bovine alpha or beta interferon. IFN-gamma from sheep, goat and buffalo, but not from pig, deer or man, are also recognised by the EIA. The bovine IFN-gamma EIA when used in conjunction with the whole blood culture system has resulted in a simple, rapid and sensitive in vitro assay for specific cell mediated immune responsiveness to M. bovis infection in cattle.     

In vitro Evaluation of the Quality and Fertilizing Capacity of Boar Semen Frozen in 0.25 ml straws

Twenty-two boar ejaculates were frozen in 0.25 ml straws using a controlled cooling rate, then evaluated in vitro in order to assess: (i) the extent to which a range of semen evaluation parameters accurately characterize sperm quality, (ii) the value of quality assessment in the characterization of long-term sperm survival and fertility and (iii) the suitability of the cryopreservation protocol used for yielding semen with good quality and fertilizing capacity. Motility with or without caffeine, plasma membrane integrity (PMI) evaluated with both propidium iodide (PI) and Hoechst 33258, and acrosome morphology were studied, the ejaculates being then classified into five quality groups. A thermoresistance test and a homologous in vitro fertilization test were applied to selected ejaculates of these groups. Caffeine-stimulated motility and PMI evaluated with PI provided better estimations of semen quality than the other tests of motility, PMI, or acrosome morphology, but this quality assessment could not reveal differences in fertilizing capacity or thermoresistance among ejaculates. Over 43% spermatozoa survived cryopreservation in 19 of the 22 ejaculates, with inter-boar and inter-ejaculate variability in the freezing success being observed. The fertilizing capacity, however, was seriously affected by the process regardless of the semen quality. It is concluded that caffeine-stimulated motility and PMI evaluated with PI give accurate information on sperm quality, but important aspects to the valuation of semen such as thermoresistance and fertilizing capacity are not revealed by this quality study. Moreover, the approach of selecting suitable protocols of cryopreservation does not appear to be sufficient for guaranteeing systematically good quality and fertilizing capacity in the frozen-thawed semen.     

Photoprotection of Beauveria bassiana: Testing simple formulations for control of the coffee berry borer

The entomopathogenic fungus, Beauveria bassiana is considered to be one of the few natural enemies available for use against the coffee berry borer. In an attempt to enhance the efficacy of this pathogen, a range of concentrations of 22 substances was tested in simple laboratory tests using natural sunlight or a UV light source. Unprotected B. bassiana spores were almost completely inactivated by exposure to 60 min of direct sunlight or 20 s of UV light of 302 nm wavelength. Seven of the 22 substances tested showed little or no photoprotective properties and eight of the substances appeared directly detrimental to spore germination. Of the remainder, sucrose, yeast, yeast extract, uric acid, casein, and molasses had limited photoprotective properties. The most effective substances tested were egg albumen and skimmed milk powder which could extend the persistence of B. bassiana spores by a factor of almost three. A mixture of 3% (w/v) albumen and 4% (w/v) milk powder gave the highest degree of spore protection per unit cost. Young coffee plants sprayed with this mixture did not suffer any significant phytotoxic effects. A field trial, involving two applications of spores with or without the milk and albumen mixture, failed to show that improved spore persistence resulted in increased coffee berry borer control. Very lowlevels of pest infestation observed in field plots together with unusual, unfavourable weather conditions may have accounted for this unexpected result.     

The edible fruit species in Mexico

In Mexico 762 edible fruit species are presently distributed into 288 genera belonging to 87 botanical families. There are 113 cultivated fruit tree species, of which 53 are native and 60 have been introduced; there are also 649 non-cultivated or underutilized species, of which 88 of them were introduced and 554 are native ones. The best representative botanical families are: Anacardiaceae (16 spp.); Anonaceae (19 spp.); Arecaceae (20 spp.); Boraginaceae (10 spp.); Cactaceae (88 spp.); Canabaceae (11 spp.); Caricaceae (9 spp.); Ebenaceae (15 spp.); Ericaceae (22 spp.); Fabaceae (37 spp.); Grossulariaceae (21 spp.); Juglandaceae (15 spp.); Lauraceae (18 spp.); Malpighiaceae (11 spp.); Malvaceae (12 spp.); Melastomataceae (13 spp.); Moraceae (20 spp.); Myrtaceae (29 spp.); Passifloraceae (9 spp.); Pinaceae (12 spp.); Rosaceae (88 spp.); Rubiaceae (9 spp.); Rutaceae (28 spp.); Sapotaceae (24 spp.); Solanaceae (23 spp.); and Vitaceae (24 spp.). This taxonomic richness is related to neartic and neotropical environments, and marked by the human groups present throughout the history of the country. The in situ conservation is recommended in southern, eastern, northeastern and northwestern regions of the country. The ex situ conservation needs consider species with non-orthodox seeds. Probably a few wild edible species need to be grown for commercial cultivation but it is not a rule for all.     

Quantification of Sexual Steroid Hormones in Faeces of Iberian Wolf (Canis lupus signatus): A Non‐invasive Sex Typing Method

The determination of gender in wild animals is essential for behavioural and ecological studies, and also for conservation. The objectives of this study were (i) the determination of gender in faecal samples of Iberian wolf based on the differential concentrations of sexual steroid hormones (SSH) and (ii) to analyse the profiles of SSH in males and females (considering the gender determination carried out previously) during the non‐reproductive and reproductive periods. The quantification of androgens (testosterone, T), progestin (progesterone, P) and oestrogen (oestradiol, E) was conducted by means of enzyme immunoassay. The k‐means conglomerate analysis showed that the 59 faecal samples grouped into three different conglomerates, considering SSH levels. Groups 1 and 2 showed higher levels of T than group 3. Therefore, the faecal samples included in groups 1 and 2 (17 samples) corresponded to males and those of group 3 (42 samples) to females. The levels of T + P + E and T/P were higher in the group of males than in the group of females. The results of this study also showed that levels of T in males were higher during the reproductive period than in the non‐reproductive period. However, the concentrations of P and E turned out to be higher during the non‐reproductive season. In females, the levels of the three hormones (T, P and E) were higher during the reproductive period.     

Seasonality of Reproduction in Mammals: Intimate Regulatory Mechanisms and Practical Implications

Farm mammals generally express seasonal variations in their production traits, thus inducing changing availability of fresh derived animal products (meat, milk and cheese) or performances (horses). This is due to a more or less marked seasonal birth distribution in sheep and goats, in horses but not cattle. Birth peak occurs at the end of winter-early spring, the most favourable period for the progeny to survive. Most species show seasonal variations in their ovulation frequency (presence or absence of ovulation), spermatogenic activity (from moderate decrease to complete absence of sperm production), gamete quality (variations in fertilization rates and embryo survival), and also sexual behaviour. The intimate mechanism involved is a complex combination of endogenous circannual rhythm driven and synchronized by light and melatonin. Profound and long-term neuroendocrine changes involving different neuromediator systems were described to play a role in these processes. In most species artificial photoperiodic treatments consisting of extra-light during natural short days (in sheep and goats and mares) or melatonin during long days (in sheep and goats) are extensively used to either adjust the breeding season to animal producer needs and/or to completely overcome seasonal variations of sperm production in artificial insemination centres. Pure light treatments (without melatonin), especially when applied in open barns, could be considered as non-invasive ones which fully respect animal welfare. Genetic selection could be one of the future ways to decrease seasonality in sheep and goats.