Method of determination of nitrosamines in sausages by CO2 supercritical fluid extraction (SFE) and micellar electrokinetic chromatography (MEKC)

In this paper, the use of supercritical fluid extraction (SFE) and micellar electrokinetic capillary chromatography (MEKC) is proposed for the complete analysis of volatile nitrosamines in sausages. The extraction fluid used was CO2 and variables such as density, temperature of thimbles, extraction time, modifier, fluid flow, and kind of traps were investigated. Several experiments were carried out to obtain the most favorable conditions for analysis of volatile nitrosamines in sausages. The recoveries ranged from 21 to 82% for the five nitrosamines studied. The optimal condition of extraction was 0.2 g of sample fortified with 10 mg/kg, using dynamic extraction during 20 min and with adsorbent Florisil in the trap. The solvent selected for the elution of the analytes was methanol.     

Chemical Composition of Green Tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis>) Infusions Commercialized in Portugal

To evaluate the potential benefits and risks associated with tea consumption it is important to identify the constituents of this beverage. Levels of some minerals, caffeine and catechins in green tea samples commercialized in Portugal were evaluated. Potassium is the metal present in larger amount (92–151 mg/l). The content of sodium, calcium, fluoride, aluminium, manganese and iron were 35–69, 1.9–3.5, 0.80–2.0, 1.0–2.2, 0.52–1.9, 0.020–0.128 mg/l, respectively. Chromium and selenium were not detected. The resulting data showed considerable variability in catechins content. The levels of epigallocatechin gallate (EGCG) ranged from 117 to 442 mg/l, epicatechin 3-gallate (EGC) from 203 to 471 mg/l, epigallocatechin (ECG) from 16.9 to 150 mg/l, epicatechin (EC) from 25 to 81 mg/l and catechin (C) from 9.03 to 115 mg/l. Caffeine contents in the green tea infusions studied were between 141–338 mg/l. Green tea infusions provide significant amounts of catechins and could be an important source of some minerals.     

Short-term baths with essential oils of Lippia sidoides,Ocimum gratissimum and Zingiber officinale influence blood parameters and survival of tambaqui (Colossoma macropomum) after infection with Aeromonas hydrophila

This study evaluated the effect of short-term baths with the essential oils (EOs) of Lippia sidoides, Ocimum gratissimum and Zingiber officinale on the survival, haematological, biochemical and histopathological parameters of native fish tambaqui (Colossoma macropomum), challenged with Aeromonas hydrophila. After challenge, the fish infected with A. hydrophila received therapeutic baths of 60 min, for five consecutive days. Treatments were as follows: challenged and not treated; challenged and treated with 10 mg L^?1 of gentamicin; challenged and treated with EO of L. sidoides (2.5 and 5.0 mg L^?1); challenged and treated with EO of O. gratissimum (5.0 and 10.0 mg L^?1); challenged and treated with Z. officinale EO (5.0 and 10.0 mg L^?1). The highest survival rates were 89.5% (fish treated with 5.0 mg L^?1 of O. gratissimum EO) and 75.0% (fish treated with 5.0 mg L^?1 of L. sidoides EO). Reductions in the haematocrit percentage, haemoglobin and erythrocyte number were observed in fish treated with L. sidoides and Z. officinale EOs compared with control fish. Mild to moderate damage to the liver tissue of fish after challenge and EO treatments were observed. It is suggested that baths containing O. gratissimum and L. sidoides EOs consist in a viable treatment to increase survival rates of C. macropomum infected with A. hydrophila.     

Prevalence and risk factors associated with goat gastrointestinal helminthiasis in the Sertão region of Paraíba State,Brazil

Gastrointestinal helminthiasis represents an obstacle to goat raising, causing severe damage to herds such as growth retardation, weight loss, and even death. In this study, we aimed to determine the prevalence and risk factors associated to goat gastrointestinal helminthiasis in the Sertão region of Paraíba State, Brazil. A total of 256 goats from 54 farms were systematically sampled. Blood and fecal samples were collected from each animal for egg per gram (EPG), larval culture, and packed cell volume (PCV) analyses. We found that 79.3 % of the goats investigated were parasitized with gastrointestinal helminths. Significant correlation (p?=?0.004) was observed between the EPG and PCV of the animals studied, and it was observed that the EPG increases as the PCV decreases. In the larval culture, the most prevalent helminth was Haemonchus sp. (83.2 %). Age and sex were significant variables (p?≤?0.20) for the development of gastrointestinal helminths: 86.8 % of animals over 36 months of age and 81.7 % of females were infected. The variable type of animal exploitation was also significant, with 90.3 % (p?≤?0.20) of the animals presenting double suitability (milk and meat). The Sertão region of Paraíba State presents high prevalence of gastrointestinal helminthiasis in goats, and age and type of animal exploitation are the most relevant risk factors to the development of these parasites.     

Brazilian foot and mouth disease status and meat exportation to the European Union

The aim of this study was to define the differences between the Brazilian states that export and do not export meat to the European Union (EU) and to identify the variables that are important to meet the export requirements. Infrastructure and computerization of the control of animal transit in Brazil that impact on regional health status were evaluated and linked to other variables such as status for foot and mouth disease (FMD) and qualification to export meat to EU. Variables related to transit control of bovines implemented by the state agencies of animal health and inspection in each Brazilian state were evaluated. Using a discriminant analysis, four variables were selected that explained the variation between Brazilian states that were “free” and “not free” of FMD while another four were selected to explain the variation between the zones “approved” and “not approved” to export meat to the EU, including number of official veterinarians, total transit of bovines and buffaloes, total number of animal transit certificates issued for bovine and buffaloes at the state or zone level, and total number of municipalities in the state or zone. It was possible to correctly discriminate between “free” and “not free” FMD states or zones. Variables related to animal transit are important in assessing the state for the classification of animal health situation and for EU approval for the exportation of meat.     

First report of Enterocytozoon bieneusi from dairy cattle in Argentina

Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.     

Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.     

Epidemiological Analysis of Leptospira spp. Infection in Equids from the Brejo Paraibano Microregion of Brazil

Leptospirosis is a zoonotic disease that occurs in humans and several animal species, including cattle, pigs, and horses, and it has a worldwide distribution. In equids, leptospirosis manifests through recurrent uveitis and abortion and other reproductive disorders. The objective of the present study was to investigate the epidemiological situation of Leptospira spp. infection in equids from the Brejo Paraibano microregion in the northeast of Brazil, that is, to assess the prevalence of and identify the major risk factors for infection. For this purpose, blood samples were taken from 257 equids (204 horses, 46 mules, and 7 donkeys), and a microscopic agglutination test (MAT) was used as serological analysis, with 24 serovars of Leptospira spp. as antigens. Antibodies were observed in 16.2% of horses, 13.0% of mules, and 28.6% of donkeys. Animals between the ages of 2.5 and 11 years had a risk of infection 0.4 times lower than those younger than 2.5 years old (odds ratio, 0.4; 95% confidence interval, 0.18-0.90). Further studies are needed to identify potential wild reservoirs and the main routes of infections.     

Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

Faecal nitrogen excretion as an approach to estimate forage intake of wethers

Data from twenty‐two digestibility trials were compiled to examine the relationship between faecal N concentration and organic matter (OM) digestibility (OMD), and between faecal N excretion and OM intake (OMI) by wethers fed tropical or temperate forages alone or with supplements. Data set was grouped by diet type as follows: only tropical grass (n = 204), only temperate grass (n = 160), tropical grass plus supplement (n = 216), temperate grass plus supplement (n = 48), tropical grass plus tropical legume (n = 60) and temperate grass with ruminal infusion of tannins (n = 16). Positive correlation between OMD and either total faecal N concentration (Nfc, % of OM) or metabolic faecal N concentration (Nmetfc, % of OM) was significant for most diet types. Exceptions were the diet that included a tropical legume, where both relationships were negative, and the diet that included tannin extract, where the correlation between OMD and Nfc was not significant. Pearson correlation and linear regressions between OM intake (OMI, g/day) and faecal N excretion (Nf, g/day) were significant for all diet types. When OMI was estimated from the OM faecal excretion and Nfc‐based OMD values, the linear comparison between observed and estimated OMI values showed intercept different from 0 and slope different from 1. When OMI was estimated using the Nf‐based linear regressions, the linear comparison between observed and estimated OMI values showed neither intercept different from 0 nor slope different from 1. Both linear comparisons showed similar R² values (i.e. 0.78 vs. 0.79). In conclusion, linear equations are suitable for directly estimating OM intake by wethers, fed only forage or forage plus supplements, from the amount of N excreted in faeces. The use of this approach in experiments with grazing wethers has the advantage of accounting for individual variations in diet selection and digestion processes and precludes the use of techniques to estimate forage digestibility.