Claw disorders as iceberg indicators of cattle welfare: Evidence-based on production system, severity, and associations with final muscle pH.

This study presents a novel approach to use claw disorders in cattle as a retrospective welfare indicator characterized at the abattoir. A total of 1040 cattle (2080 front and back left claws) were analyzed from 143 batches, originating from feedlots, free-range, and dairy systems. Our results indicate that abnormal claw shapes (>55%) and fissures of the claw wall (>25%) had the highest prevalence, regardless of the system of origin. For the seven types of lesions monitored, numerous associations were found between lesions in the front and rear limbs typical of each production system. Ultimate meat pH was higher in animals with white line disease and skin wounds in feedlot and free-range cattle. We conclude that claw disorders can be used as an iceberg indicator to provide valuable information about animal fitness, and the ability to cope with the husbandry and pre-slaughter environment. These indicators can be used to improve the level of welfare of the animals.

Beyond scoring systems: usefulness of morphometry considering demographic variables, to evaluate neck and overall obesity in Andalusian horses.

Morphometry has proven to be a useful tool, both for the clinician and horse owners, for evaluating the body condition in equids due to its objectivity, easiness and capacity for detection of important metabolic disturbances. However, limited information is available on the use of morphometric ratios to characterize regional and overall adiposity and much less about their application in different genders, ages and horses with different levels of obesity. The objectives were to evaluate body and neck absolute measurements and ratios; factors affecting them such as the influence of gender, age, appearance of the neck crest and overall body condition and; relationships among these measurements. A total of 154 Andalusian horses classified according to their gender, age, body score status and cresty neck condition were evaluated in this cross-sectional study. Two evaluators assigned a body condition score (BCS, 1 to 9) and a cresty neck score (CNS, 0 to 5) to each horse. Horses were divided into males and females; young (2 to 5 years) and adults (6 to 15 years); obese (BCS⩾7) and non-obese (BCS<7); cresty neck (CNS⩾3) and non-cresty neck horses (CNS<3). Morphometric measurements (cm) included were: height at the withers (HW); body length (BL), girth (GC) and waist (WC) circumferences; neck length (NL); three neck circumferences (NCs), over the first (NC25%), the second (NC50%) and the third part (NC75%) of the NL and neck crest height (NCH). These measurements were also used to calculate the following ratios: GC : HW, WC : HW, GC : BL, WC : BL, NC25% : HW, NC50% : HW, NC75% : HW, NC25% : BL, NC50% : BL, NC75% : BL, NC25% : NL, NC50% : NL, NC75% : NL, NC25% : NCH, NC50% : NCH and NC75% : NCH. The results showed that most of the absolute measurements and ratios were greater than those described in other light breeds. In addition, most neck ratios were higher (P<0.050) in males than in females, however, all body ratios were greater (P<0.001) in females. Among the absolute measurements, WC in obese horses and NC25% and NC75% in cresty neck horses highlighted as higher. Either GC : HW or WC : HW and NC75% : BL were alternative surrogates for the appraisal of overall and regional adiposity in Andalusians. Several interactions were observed between the gender and adiposity scoring systems affecting the morphometric evaluation. This study establishes absolute morphometric measurements and ratios in Andalusian horses. It also highlights the variability of morphometric values and how the outcome of these can be influenced by demographic variables and the breed analyzed. Further studies are necessary to set morphometric reference values in other breeds.

Comparison of autologous bone marrow and adipose tissue derived mesenchymal stem cells, and platelet rich plasma, for treating surgically induced lesions of the equine superficial digital flexor tendon.

Several therapies have been investigated for equine tendinopathies, but satisfactory long term results have not been achieved consistently and a better understanding of the healing mechanism elicited by regenerative therapies is needed. The aim of this study was to assess the separate effects of autologous bone marrow (BM) and adipose tissue (AT) derived mesenchymal stem cells (MSCs), and platelet rich plasma (PRP), for treating lesions induced in the superficial digital flexor tendon (SDFT) of horses. Lesions were created surgically in both SDFTs of the forelimbs of 12 horses and were treated with BM-MSCs (six tendons), AT-MSCs (six tendons) or PRP (six tendons). The remaining six tendons received lactated Ringer's solution as control. Serial ultrasound assessment was performed prior to treatment and at 2, 6, 10, 20 and 45 weeks post-treatment. At 45 weeks, histopathology and gene expression analyses were performed. At week 6, the ultrasound echogenicity score in tendons treated with BM-MSCs suggested earlier improvement, whilst all treatment groups reached the same level at week 10, which was superior to the control group. Collagen orientation scores on histological examination suggested a better outcome in treated tendons. Gene expression was indicative of better tissue regeneration after all treatments, especially for BM-MSCs, as suggested by upregulation of collagen type I, decorin, tenascin and matrix metalloproteinase III mRNA. Considering all findings, a clear beneficial effect was elicited by all treatments compared with the control group. Although differences between treatments were relatively small, BM-MSCs resulted in a better outcome than PRP and AT-MSCs.

Spatial Epidemiology and Risk Factor Analysis of White Spot Disease in the Shrimp Farming Industry of Sinaloa, Mexico, from 2005 to 2011.

White spot disease (WSD), caused by the white spot syndrome virus, is currently one of the primary causes of mortality and economic losses in the shrimp farming industry worldwide. In Mexico, shrimp production is one of the most important primary activities generating an annual income of USD 711 million. However, WSD introduction in 1999 had a devastating impact for the Mexican shrimp industry. The aim of this study was to characterize the WSD spatio-temporal patterns and to identify the primary risk factors contributing to WSD occurrence from 2005 to 2011 in Sinaloa, Mexico. We used data collected by the 'Comité Estatal de Sanidad Acuícola de Sinaloa' from 2005 to 2011 regarding WSD outbreaks as well as environmental, production and husbandry factors at farm level. The spatio-temporal patterns of WSD were described using space-time scan statistics. The effect of 52 variables on the time to WSD outbreak occurrence was assessed using a multivariable Cox proportional hazards model. Results reveal that WSD risk and survival time were not homogeneously distributed as suggested by the significant clusters obtained using the space-time permutation model and the space-time exponential model, respectively. The Cox model revealed that the first production cycle [hazard ratio (HR) = 11.31], changes from 1 to 1.4°C of temperature oscillation caused by 'El Niño'/'La Niña' events (HR = 1.44) and high average daily growths (HR = 1.26) were significantly associated with lower survival (i.e. shorter time to WSD outbreak) on farm. Conversely, shrimp weight at the moment of the outbreak (HR = 0.159), changes from -0.9 to -0.5°C of temperature oscillation caused by 'El Niño'/'La Niña' events (HR = 0.540), high superficial water temperature during the pound stocking (HR = 0.823) and high (>100) number of days of culture (HR = 0.830) were factors associated with higher survival. Results are expected to inform the design of risk-based, intervention strategies to minimize the impact of WSD in Mexico.

Proposal of a production and management index (PMI) for tilapia farms.

Tilapia is one of the most important species in aquaculture; however, there is no available index to show the performance of a production unit. It is desirable to assess the productivity using indexes, such as the production and management index for shrimps and the European production efficacy factor for broilers. These indexes are based on data production: growth, survival, and feed conversion of a full production cycle. Taking into account these parameters, we propose a production and management index (PMI) for tilapia that is applicable for a specific period of the production cycle. For the construction and validation of the PMI we have used production data from 8,614 monthly records of 2 tilapia farms in Huila Department (Colombia), and because of the complexity of tilapia management, different anomalous situations have been detected and then defined as exceptions. As a result, 419 records were considered extreme values because 1 or more exceptions were met. The value of the PMI varies from 0 (the worst situation) to 3.55, which reflects high variability. We have constructed a PMI for tilapia as the product of 3 elements to obtain a positive value index. Instead of classic parameters, we had to calculate an adapted version of them: the relative average daily growth, the survival (as a complementary value of the estimated monthly mortality), and a feed conversion ratio index. To assess the utility of the PMI, some comparisons were performed using records from black and red tilapia. We observed significant differences depending on tilapia strain (PMI = 1.0248 vs. PMI = 1.1661; < 0.001), age (better values for small fish), and season (PMI = 1.0847 vs. PMI = 1.1011; = 0.026). According to these results, we can conclude that the PMI could be a useful tool for tilapia farmers, despite the complexity of the calculation.

Endocrine, morphometric, and ultrasonographic characterization of neck adiposity in Andalusian horses.

Equine metabolic syndrome (EMS) can be diagnosed by hormonal measurements; however, it would be important to find simpler measurements that allow easy identification of affected or at risk individuals. In horses, the dorsal neck region is one of the most frequent anatomical sites for fat deposition and neck obesity has been linked to EMS. The aim of this study was to evaluate the association of hormonal markers of obesity (leptin) and insulin resistance (insulin) with morphometric and ultrasonographic neck measurements in Andalusian horses. Plasma leptin and insulin concentrations were measured by RIA in 127 Andalusian horses. Neck circumferences (NC) were measured at 3 equidistant locations at 25%, 50%, and 75% of neck length (NC-25%, NC-50%, and NC-75%). At the same 3 locations, subcutaneous fat thickness (SFT-25%, SFT-50%, and SFT-75%) was measured ultrasonographically. In the population under study, a tendency to adiposity was confirmed by the elevated plasma leptin levels (7.47 ± 5.03 ng/mL). However, plasma insulin concentrations (4.05 ± 3.74 µIU/mL) were within normal range in most horses. Our results indicate that NC showed significant sexual dimorphism and did not correlate well with hormonal measurements. Ultrasonographic assessment of fat thickness at the base of the neck (SFT-75%) was significantly correlated with both plasma leptin and insulin and did not show differences between males and females. Thus, in the search for a single objective parameter which can be used in large populations, SFT-75% is a potential candidate and may be a meaningful parameter to predict EMS.

THE COMBINED USE OF HONEY, GARLIC (ALLIUM SATIVUM L.) AND SKIMMED MILK AS AN EXTENDER FOR CHILLING SHEEP SEMEN.

BACKGROUND: Sugars are the energetic source for sperm to maintain the metabolic process, and the antibiotics slow down sperm degradation. OBJECTIVE: To study the effects of rosemary honey as energy source and cryoprotectant in combination with garlic as a natural antibiotic on the quality of ram spermatozoa upon cooling. MATERIALS AND METHODS: The ejaculates from three rams were evaluated at different times during cooling to determine its post-dilution quality. RESULTS: Glycerol and dimethylformamide in conjunction with honey and garlic significantly improve the survival of spermatozoa. CONCLUSION: The addition of honey and garlic reduces sperm deterioration when stored at 4 degree C.

Freeze-dried dog sperm: Dynamics of DNA integrity.

Freeze-drying (FD) has been proposed as an alternative method to preserve spermatozoa. During the FD procedure, sperm DNA might become damaged by both freezing and drying stresses caused by the endonucleases, the oxidative stress and the storage conditions. We examined the DNA integrity of dog sperm freeze-dried with two kinds of chelating agents in FD buffers and storage at two different temperatures. Ejaculated sperm from four dogs were suspended in basic medium (10 mM Tris-HCl buffer+50 mM NaCl) supplemented with 50 mM EGTA or with 50 mM EDTA and then freeze-dried. Sperm samples were stored at 4°C as room temperature, and the analysis of DNA damage was performed after a month and 5 months of storage using a Sperm Chromatin Dispersion test. We found four different sperm populations according to the size of the halos around the sperm head: (1) absent halo, (2) <6 µm, (3) 6-10 µm, (4) >10 µm. All of them coexisted in each freeze-dried dog semen samples and differed significantly among different treatments. The highest percentage of spermatozoa with halo >10 µm was obtained when the semen samples were freeze-dried in EDTA medium and stored at room temperature for five months. Results suggested that both, the kind of chelating agent as well as storage temperature and period, influenced DNA integrity of freeze-dried dog sperm.

Sage (Salvia officinalis) and fennel (Foeniculum vulgare) improve cryopreserved boar epididymal semen quality study.

The aim of this study was to evaluate the effect of fennel and sage extracts and the influence of the egg yolk source (fresh or pasteurized) on the success of freezing boar epididymal spermatozoa. In experiment 1, epididymal sperm was recovered by flushing and cryopreserved in a lactose-egg yolk solution supplemented with various concentrations (10, 5 and 2.5 g/L) of sage or fennel. Sperm quality was evaluated (motility, viability, HOST and acrosome integrity) at 0 h and 2 h after thawing. Fennel 10 g/L and sage 5 g/L and control (no extracts) were selected for experiment 2 which also compared fresh or pasteurized egg yolk in the freezing extender and measured DNA integrity of the frozen sperm. Results showed that the interaction between fennel and sage antioxidants with fresh egg yolk significantly improved post thaw sperm quality and protected boar epididymal spermatozoa from cryopreservation damage as a result of oxidative stress.

Epidemiological survey of brucellosis in sheep and goats in selected pastoral and agro-pastoral lowlands of Ethiopia.

An epidemiological survey was conducted in pastoral regions of Ethiopia to investigate the distribution of brucellosis in sheep and goats. Between November 2004 and December 2007, a total of 6,201 serum samples were collected from 67 randomly selected peasant associations, 25 districts and eight pastoral zones of Ethiopia. The Rose Bengal plate test (RBPT) and complement fixation test were used in series. Samples for bacteriology were collected from three export abattoirs, where 285 goats were randomly selected and tested by RBPTthree days before slaughter. Tissue samples were collected from 14 strongly positive goats and cultured in dextrose agar and Brucella agar base. To confirm and subtype the isolates, staining, biochemical tests and polymerase chain reaction were used. The overall standardised seroprevalence of brucellosis was 1.9%, ranging from 0.07% in Jijiga zone to 3.3% in Borena zone. There was statistically significant variation among the studied regions, zones, districts and peasant associations (p < 0.05). Male goats and sheep were twice as likely to test positive as females (relative risk [RRJ: 2.04; 95% confidence interval [CI]:1.7-3.4; x2 = 21.05, p < 0.05). Adults (older than 1.5 years) were three times more likely to test positive than younger animals (RR: 2.76; 95% CI: 1.14-6.73; chi2 = 5.18, p < 0.05). Goats were around four times more likely to be infected than sheep (RR: 3.8; 95% CI: 2.4-6.1; chi2 = 36.99, p < 0.05). Brucella melitensis was isolated from 2 of the 14 samples analysed. The widespread distribution of brucellosis in goats and sheep in these areas justifies the use of control measures to minimise the economic losses and public health hazards.

Antioxidant effect of lemon balm (Melissa officinalis) and mate tea (Ilex paraguensys) on quality, lipid peroxidation and DNA oxidation of cryopreserved boar epididymal spermatozoa.

In this study, we investigated the protective ability of the addition of two antioxidant herb extracts, mate tea and lemon balm, on boar epididymal frozen-thawed spermatozoa quality. Testes from mature boars were collected at local slaughterhouse, and sperm samples from epididymis were recovered by flushing. Spermatozoa were cryopreserved in lactose-egg yolk buffer supplemented with various concentrations of lemon balm and mate tea (0, 2.5, 5 and 10 g l(-1) ) using the straw-freezing procedure. Motion parameters, acrosome and plasma membrane integrity, lipoperoxidation levels and DNA oxidative damage (8-hydroxy-2'-deoxyguanosine base lesion) were evaluated. There were no differences among experimental groups with regard to motility characteristics, viability, acrosome and plasma membrane integrity; however, the highest concentration of lemon balm produced significant (P < 0.05) improvement in curvilinear trajectory, straightness and amplitude of lateral head displacement after thawing. The supplementation of freezing extender with mate tea and lemon balm reduced sperm lipid membrane peroxidation, and only mate tea protected DNA against oxidative damage during cryopreservation at 120 min post-thawing (P < 0.05). Mate tea experimental extender at concentration of 10 g l(-1) showed the lowest percentage of sperm oxidised DNA and malondialdehyde generation; thus, mate tea is a potential candidate such as antioxidant compound on boar sperm cryopreservation medium.

Effect of different disaccharides on the integrity and fertilising ability of freeze-dried boar spermatozoa: a preliminary study.

Freeze-drying spermatozoa is a developing technique that facilitates semen storage and transport. However, freeze-dried sperm exhibits impaired DNA integrity, which is associated with reduced fertilizing ability. Boar spermatozoa were freeze-dried in three different freeze-drying EDTA buffers with trehalose (75mM) and lactose (75mM) (EDTA-TL), (2) with sucrose (75mM) and lactose (75mM) (EDTA-SL) or just lactose (150mM) (EDTA-LL) using two freeze-drying protocols. In experiment 1 a one-step protocol was used and in experiment 2 a two-steps protocol was used. Spermatozoa were stored in1.5 mL cryo-tubes and 1.5 mL glass ampules at both 16 degree C and 25 degree C for 1 month. Successfully freeze-dried spermatozoa were stained with acridine-orange to assess chromatin stability. Freeze-drying was most successful when the 2-step protocol was used (experiment 2). Chromatin stability was greater in samples stored in glass ampules compared to cryo tubes. Chromatin stability was also greater in samples freeze-dried in EDTA-LL compared to EDTA-SL or EDTA-TL buffers. Spermatozoa freeze-dried in EDTA-LL and stored for 14 and 28 days at either 16 degree C or 25 degree C were utilized for ICSI. Two pronuclear formation wasgreatest using spermatozoa stored at 25 degree C (69.23%) and for 28 days (50%). Although 16 degree C spermatozoa samples had better stable chromatin, 25 degree C spermatozoa samples offered better two pronuclear formation results. In conclusion, boar spermatozoa freeze-dried using media containing disaccharides exhibit high chromatin stability and are able to fertilise oocytes following ICSI. Disaccharides may therefore advance the development of freeze-drying techniques for spermatozoa enabling ease of sperm storage and transportation.

Effect of pasteurized egg and Rosmarinus officinalis supplementation on quality of cryopreserved ram semen.

The aim was to assess the in vitro effect of pasteurized egg (PE) and rosemary (Rosmarinus officinalis) on frozen-thawed ram semen. Ejaculates from three mature rams of the Rasa Aragonesa breed were cryopreserved using a 2-step dilution method (Fraction 1: F1; Fraction 2: F2). In Experiment 1, semen was frozen in egg yolk (EY) or PE extenders. After thawing, similar results were obtained in terms of total and progressive motility, viability, hypo-osmotic swelling test (HOST) and acrosome integrity after 2 h incubation. In Experiment 2, addition of rosemary to F1, F2 or both fractions to EY extenders was evaluated. Rosemary in F1 decreased progressive motility (p = 0.013) after 2 h incubation. Finally, PE can be used as a substitute for EY to reduce hygienic risks in extenders and is easier to standardize. Supplementation of EY extender with rosemary in F1 reduced progressive motility. Rosemary supplementation in F2 does not affect semen quality.

Determination of ovulation time in sows based on skin temperature and genital electrical resistance changes.

Different physical and physiological parameters may be used to determine ovulation time in sows. In the present study, we analysed the ear and vulvar skin temperature fluctuations, and the changes in genital electrical resistance, at a distance of 4, 8 and 12 cm from the vulva during oestrus in order to predict the time of ovulation. Multiparous sows were checked by transrectal real-time ultrasonography and luteinising hormone (LH) plasma concentration was determined. Temperature was measured using a thermoprecision infrared thermometer, and the electrical resistance was measured with a commercial resistance probe. All measurements were carried out every 12 hours from one day after the weaning to three days after oestrus onset. Skin temperature showed significant difference around periovulatory period. The electrical resistance at 4 cm from the vulva showed marked changes during oestrus, which were different from those described at 8 and 12 cm from the vulva. At 12 hours before ovulation time, skin temperature decreased significantly, and negative correlation (P<0.05) was found between vulvar skin temperature and vaginal resistance. There was no relationship between skin temperature, electrical resistance and LH plasma concentration. The measurement of several physiological traits may provide more accurate predictions of the moment of ovulation.

Crystallisation pattern of vestibular mucus and its relation to vestibular electrical resistance in cycling sow.

Changes in the genital mucus around the oestrus are used by different diagnostic methods to determine optimal fertilisation time. In the current study, the authors evaluated the different arborisation patterns found in vestibular mucus, and also established its relationship with vestibular resistance changes during oestrus. Thirty multiparous sows were checked by transrectal ultrasonography to determine ovulation time every 12 hours. Vestibular resistance was measured with a commercial resistance probe, and vestibular mucus ferning was also evaluated every 12 hours during the oestrus. Significant changes (P < 0.05) in vestibular resistance were detected, registering high variation among individuals. Maximum resistance data was reached between 12 and 24 hours after ovulation time in 83 per cent of the sows. Crystallisation samples were classified into three different patterns according to the fern-like crystal degree. Arborisation peak occurred from 48 to 36 hours before the moment of ovulation, when vestibular resistance values increased gradually. In the optimal insemination moment, vestibular resistance increased significantly (P < 0.05) and vestibular mucus showed a low crystallisation pattern (P < 0.05). Combining several methods to measure genital mucus changes may predict the ovulation time and the best insemination moment.

Persistence of sucrose fermenting and nonfermenting vibrios in tissues of Manila clam species, Ruditapes philippinarum, depurated in seawater at two different temperatures.

Effective post-harvest treatment to eliminate toxigenic vibrios is an important measure to reduce risk associated with seafood consumption. In the present study, we examined whether natural populations of sucrose nonfermenting (SNFV) and sucrose fermenting vibrios (SFV) persisted in Ruditapes philippinarum, after depuration at two different temperatures. Two experiments (one in Spring and one in Summer) were performed. Clams were depurated in duplicate tanks in an open-circuit seawater system using filtration and ultraviolet treatment. Clams were sampled daily (9 days), individually processed and inoculated on thiosulphate-citrate-bile salts-sucrose (TCBS) agar plates to estimate the density of SNFV and SFV. Results showed that depuration was less effective in Summer than in Spring, for SNFV and SFV. Despite an initial lower presence in the Summer experiment, clearance of SNFV was less efficient than in Spring, with 22.2% of clams presenting SNFV on day 8 and without significant changes on day 9. When compared, daily proportion of presence of SNFV and SFV in Spring, it was observed that SFV were more frequent than SNFV on days 8 and 9. In Summer these differences were significant on days 7-9. Present study demonstrated that SNFV (involved in food poisoning) are purged, at any temperature, faster than SFV.

Identification and characterization of lactic acid bacteria isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), with inhibitory activity against Lactococcus garvieae.

A study was conducted to evaluate the probiotic properties of endogenous rainbow trout microbiota against pathogenic Lactococcus garvieae. A total of 335 bacterial strains were isolated from rainbow trout and screened for antagonistic activity against L. garvieae using an agar spot assay. Antagonistic strains were grouped by PCR amplification of repetitive bacterial DNA elements (rep-PCR) and identified by 16S rRNA gene sequence analysis. The results revealed that the antagonistic strains belonged to the genera Lactobacillus, Lactococcus and Leuconostoc. Further probiotic characteristics, such as specific growth rate, doubling time, resistance to biological barriers, antibiotic resistance, hydrophobicity and production of antimicrobial substances, were also studied. These strains were able to survive low pH and high bile concentrations, showed good adherence characteristics and a broad spectrum of antibiotic resistance. The antagonistic efficacy was maintained after sterile filtration and was sensitive to proteinase K, indicating that proteinaceous extracellular inhibitory compounds were at least partially responsible for pathogen antagonism. Based on these results, these strains should be further studied to explore their probiotic effects in challenge experiments in vivo. This study shows clear evidence that the indigenous trout-associated microbiota may provide a defensive barrier against L. garvieae.

Experimental uptake and retention of pathogenic and nonpathogenic Vibrio parahaemolyticus in two species of clams: Ruditapes decussatus and Ruditapes philippinarum.

AIMS: To describe uptake and retention of pathogenic and nonpathogenic Vibrio parahaemolyticus in Ruditapes decussatus and Ruditapes philippinarum. METHODS AND RESULTS: Ruditapes decussatus accumulated greater concentrations of pathogenic and nonpathogenic V. parahaemolyticus than R. philippinarum. These concentrations decreased earlier in R. decussatus. Nonpathogenic V. parahaemolyticus reached higher concentrations (approx. 1 log CFU g⁻¹ in tissues of R. decussatus and more than 1 log CFU g⁻¹ in R. philippinarum) than pathogenic V. parahaemolyticus at similar times. It also persisted longer in both species of clams (72 h in R. decussatus and 96 h in R. philippinarum), while pathogenic V. parahaemolyticus persisted 48 h in R. decussatus and 72 h in R. philippinarum. CONCLUSIONS: Ruditapes decussatus incorporated both isolates of V. parahaemolyticus faster than R. philippinarum and it eliminated both isolates earlier than R. philippinarum. Under same conditions, nonpathogenic V. parahaemolyticus might survive better than pathogenic V. parahaemolyticus within both species of clam. SIGNIFICANCE AND IMPACT OF THE STUDY: Pathogenic V. parahaemolyticus is an important cause of foodborne illnesses. This study shows it may be possible to use nonpathogenic V. parahaemolyticus to perform experimentation to evaluate bacterial evolution in clams, developing an in vivo model useful for risk analysis.

Anti-oxidant supplementation improves boar sperm characteristics and fertility after cryopreservation: comparison between cysteine and rosemary (Rosmarinus officinalis).

Anti-oxidants partially ameliorated the detrimental effects of reactive oxidative substances produced during cryopreservation. The objective of the study was to determine the effect of anti-oxidant addition to the freezing extender on boar semen qualities and fertility capacity. Ejaculates were collected from a previously selected boar and semen samples were processed using the straw freezing procedure. In experiment 1, semen samples were cryopreserved in lactose-egg yolk solution supplemented with various concentrations of cysteine (0, 5 and 10mM) to determinate a cysteine concentration capable of producing a protective effect during cryopreservation. Semen quality (total motility, progressive motility, viability, acrosome integrity and hypoosmotic swelling test) was evaluated after freezing and thawing and then every hour for 3h. In experiment 2, ejaculates were cryopreserved with lactose-egg yolk extender with or without the following anti-oxidants: cysteine, rosemary (Rosmarinus officinalis) and cysteine plus rosemary. Semen quality was evaluated. In the experiment 3, fertility capacity of semen frozen in anti-oxidant supplementation extenders was examined in vitro. A total of 2232 oocytes were in vitro matured and inseminated with frozen-thawed sperm. In summary: (i) the effective concentration of cysteine in freezing extender was 10mM; (ii) the addition of exogenous rosemary or cysteine to the freezing extender positively affected post-thawed viability and acrosome integrity. Only rosemary supplementation improved total motility at 3h and progressive motility at any time; (iii) the inclusion of rosemary into the extender was effective in penetration and cleavage rate and also in the efficiency of the fertilization system.

Comparing sugar type supplementation for cryopreservation of boar semen in egg yolk based extender.

Cryopreservation of boar semen is still considered suboptimal due to lower fertility when compared to fresh semen. The aim of this study was to evaluate the effects of the addition of different sugars (lactose, trehalose and glucose) on boar spermatozoa cryopreserved in an egg yolk based extender. Ejaculates were collected from a boar previously selected and semen samples were processed using the straw freezing procedure. In experiment 1, subsamples of semen were frozen in three different extenders: recommended lactose egg yolk extender (LEY); trehalose egg yolk extender (TEY) and glucose egg yolk extender (GEY). Sperm quality was assessed for motility, viability, acrosome integrity and hypoosmotic swelling test response upon collection, after freezing and thawing and then every hour for 3h. Results showed that total motility at 1 and 3h, progressive motility at 3h, positive hypoosmotic response at 2 and 3h and acrosome integrity at all times were significantly improved when trehalose was added to the extender. In experiment 2, sugar influence was also demonstrated in vitro fertilization. A total of 1691 oocytes were in vitro matured and inseminated with frozen-thawed sperm at 2000:1 sperm:oocyte ratio and coincubated for 6h. Presumptive zygotes were cultured in NCSU-23 medium to assess fertilization parameters and embryo development. Both penetration and monospermy rates were significantly higher for trehalose frozen semen. A significant increase was observed in efficiency and blastocyst formation rates from TEY to the other groups. Our results demonstrated that trehalose extender enhances spermatozoa viability and its in vitro fertilization parameters in boar ejaculates with good sperm freezability. Further studies are necessary to assess the impact of sugars on the entire population.