Anthelmintic resistance of gastrointestinal nematodes in sheep grazing in irrigated and dry areas in the semiarid region of northeastern Brazil.

This study aimed to determine the AR of gastrointestinal nematodes (GIN) to commercial drugs in sheep flocks naturally infected, grazing in irrigated (IA) and dry (DA) areas of the semiarid region in northeastern Brazil. Fecal egg count reduction tests (FECRT) were performed at 10 farms. From each flock, 36 adult sheep were selected and divided into five groups (G1 (0.08% ivermectin), G2 (10% albendazole), G3 (5% levamisole), G4 (1% moxidectin), G5 (10% closantel) and one control group, G6). All the commercial drugs were found to reduce the number of eggs per gram of feces (EPG). Resistance to ivermectin (37.1%), albendazole (52.1%), and levamisole (52.0%) was detected at all the farms, but nematodes proved to be susceptible to moxidectin (87.9%) and closantel (83.9%). The overall average efficacy of the commercial drugs was significantly higher (P < 0.05) in DA (49.2%), where moxidectin (90.4%) showed high effectiveness. The presence of the parasite Haemonchus contortus predominated at all the farms. The variables irrigated area (P = 0.002), intensive breeding (P = 0.018), uncovered enclosures (P = 0.05), cultivated (P = 0.043) and native/cultivated (P = 0.007) pastures, and rotational grazing (P = 0.013) were significantly associated with GIN infection; irrigated area (P = 0.009), semi-intensive breeding (P = 0.05), rotational grazing (P = 0.045), cultivated (P = 0.021) and native/cultivated (P = 0.04) pastures, and estimated weighing of animals (P = 0.002) were significantly associated with AR. Therefore, improved management practices and strategic deworming must be implemented to prevent the development of AR.

Antimicrobial activity of propolis extract fractions against Staphylococcus spp. isolated from goat mastitis / Atividade antimicrobiana de frações do extrato de própolis frente à Staphylococcus spp. isolados de mastite caprina

The indiscriminate use of antibiotics in the treatment of caprine mastitis causes the appearance of resistant microorganisms, besides leaving residues in milk, putting at risk to human health. In this way, propolis is an alternative in the treatment of diseases because it has antimicrobial activity, mainly because of the presence of flavonoids in its composition. The aim of this study was to evaluate the antimicrobial potential of propolis to Staphylococcus spp. Isolated from cases of goat mastitis and qualify the crude ethanoic extract by high performance liquid chromatography (HPLC). In this study, the minimum bactericidal concentration values of propolis extracts in ethanol, ethyl acetate and hexane showed that the best concentrations capable of promoting the highest mortality of the isolates of Staphylococcus spp. from mastitis in goats, were 6250, 3125 and 1562.5µg/mL, respectively. By the microplate adherence test, it was found that 20.78% isolates were not able to form biofilm, 14.70% were classified as moderate and 64.70% were weak and none as a strong biofilm producer. Propolis in its different diluents was able to affect the formation of biofilm and showed a pronounced marked antimicrobial activity against Staphylococcus spp. strains and may be indicated for use in in vivo studies.(AU)

O uso indiscriminado de antibióticos no tratamento de mastite caprina leva ao desenvolvimento de micro-organismos resistentes que poderão estar presentes em alimentos, colocando em risco a saúde humana. Dessa forma, a própolis surge como uma alternativa para o tratamento de doenças por possuir uma ação antimicrobiana, principalmente pela presença de flavonoides em sua composição. O objetivo desse estudo foi avaliar o potencial antimicrobiano da própolis frente à Staphylococcus spp. isolados de casos de mastite caprina e qualificar o extrato etanoico bruto por cromatografia líquida de alta eficiência (CLAE-DAD). Neste estudo, os valores de concentração bactericida mínima (CBM) dos extratos de própolis em álcool etílico, acetato de etila e hexano nos isolados foram de 6250, 3125 e 1562,5µg/mL, respectivamente. Pelo teste de aderência à microplacas, observou-se que 20,78% dos microorganismos, não foram capazes de formar biofilme, 14,70% foram classificados como moderados, 64,70% em fracos e nenhum como forte produtor de biofilme. A própolis em seus diferentes diluentes foi capaz de afetar a formação de biofilme e apresentou significativa atividade antimicrobiana frente a cepas de Staphylococcus spp., podendo ser indicada para utilização em estudos "in vivo".(AU)

Impact of water restriction periods on carcass traits and meat quality of feedlot lambs in the Brazilian semi-arid region.

Water restriction periods were evaluated in crossbred lambs (n = 32) distributed in one of four treatments: without water restriction, water restriction for 24, 48 and 72 h. The water restriction for 72 h reduced the water and dry matter intakes, body weight at slaughter and hot and cold carcass yields. Water restriction did not affect the weight of the carcass cuts and the chemical composition of the meat. The fatty acid EPA increased and DHA reduced with increasing water restriction period. There was an increasing linear effect for meat shear force, with less force (30.5 N/cm2) for sheep meat without water restriction and higher force (45.8 N/cm2) for those with water restriction for 72 h. The period of 24 h of water restriction was the one that promoted the highest similarities in the characteristics assessed to those in animals receiving water ad libitum. Therefore, water restriction periods should not exceed 24 h for feedlot animals in situations of severe water shortage.

Human relaxins (RLNH1, RLNH2), their receptor (RXFP1) and fetoplacental growth.

Relaxin, a systemic and placental hormone, has potential roles in fetoplacental growth. Human placenta expresses two RLN genes, RLNH1 and RLNH2 Maternal obesity is common and is associated with abnormal fetal growth. Our aims were to relate systemic and cord blood RLNH2, placental RLNs and their receptor (RXFP1) with fetoplacental growth in context of maternal body mass index, and associations with insulin-like growth factor 2 (IGF2) and vascular endothelial growth factor A (VEGFA) in the same placentas. Systemic, cord blood and placental samples were collected prior to term labor, divided by prepregnancy body mass index: underweight/normal (N = 25) and overweight/obese (N = 44). Blood RLNH2 was measured by ELISA; placental RLNH2, RLNH1, RXFP1, IGF2 and VEGFA were measured by quantitative immunohistochemistry and mRNAs were measured by quantitative reverse transcription PCR. Birthweight increased with systemic RLNH2 only in underweight/normal women (P = 0.036). Syncytiotrophoblast RLNH2 was increased in overweight/obese patients (P = 0.017) and was associated with placental weight in all subjects (P = 0.038). RLNH1 had no associations with birthweight or placental weight, but was associated with increased trophoblast and endothelial IGF2 and VEGFA, due to female fetal sex. Thus, while systemic RLNH2 may be involved in birthweight regulation in underweight/normal women, placental RLNH2 in all subjects may be involved in placental weight. A strong association of trophoblast IGF2 with birthweight and placental weight in overweight/obese women suggests its importance. However, an association of only RLNH1 with placental IGF2 and VEGFA was dependent upon female fetal sex. These results suggest that both systemic and placental RLNs may be associated with fetoplacental growth.

Influence of drinking water salinity on carcass characteristics and meat quality of Santa Inês lambs.

This study aimed to evaluate the effects of different salinity levels in drinking water on the quantitative and qualitative characteristics of lamb carcass and meat. Ram lambs (n = 32) were distributed in a completely randomized design with four levels of salinity in the drinking water (640 mg of total dissolved solids (TDS)/L of water, 3188 mg TDS/L water, 5740 mg TDS/L water, and 8326 mg TDS/L water). After slaughter, blending, gutting, and skinning the carcass, hot and biological carcass yields were obtained. Then, the carcasses were cooled at 5 °C for 24 h, and then, the morphometric measurements and the cold carcass yield were determined and the commercial cuts made. In the Longissimus lumborum muscle color, water holding capacity, cooking loss, shear force, and chemical composition were determined. The yields of hot and cold carcass (46.10 and 44.90%), as well as losses to cooling (2.40%) were not affected (P > 0.05) by the salinity levels in the water ingested by the lambs. The meat shear force was 3.47 kg/cm2 and moisture, crude protein, ether extract, and ash were 73.62, 22.77, 2.5, and 4.3%, respectively. It is possible to supply water with salinity levels of up to 8326 mg TDS/L, because it did not affect the carcass and meat characteristics of Santa Inês lambs.

Carcass and non-carcass component yields of crossbred Boer and Brazilian semiarid indigenous goats subjected to different feeding levels.

The effect of different feeding levels (ad libitum, 25 and 50 % restriction) and genotypes (½ Boer × ½ nondescript breed goats, Canindé, and Moxotó) on carcass quantitative characteristics and non-carcass components (NCC) were evaluated. Forty-five intact male goats were distributed in a 3 × 3 factorial design with five replicates. There was no effect of genotype on carcass weights and yields and retail cuts weights and yields (P > 0.05). Compared to Moxotó, ½ Boer presented better carcass conformation and higher weights (P < 0.01) and yields of viscera from gastrointestinal tract (P < 0.05), and compared to the other genotypes (P < 0.01), ½ Boer presented larger carcass compactness. Carcass weights and yields, retail cuts weights and NCC, and soft tissues yields were higher (P < 0.01) in goats fed ad libitum. The two restriction levels did not differ (P > 0.05) for these variables. There were interactions of genotype and feeding level. At ad libitum feeding, ½ Boer had higher weights of breast and shank, leg, soft tissues, and gastrointestinal viscera compared to the Moxotó (P < 0.05). The crossing of nondescript breed goats with Boer may be a strategy for increasing the efficiency of goat meat production in the Brazilian semiarid. Moreover, in times of feed scarcity, farmers may use higher feed restriction levels to keep animals, since for most of the parameters evaluated, there were no differences between the restriction levels.

Relaxin, its receptor (RXFP1), and insulin-like peptide 4 expression through gestation and in placenta accreta.

This study was designed to show whether placental relaxin (RLN), its receptor (RXFP1), or insulin-like peptide 4 (INSL4) might have altered expression in patients with placenta accreta. The baseline expression of their genes through gestation (n = 34) was quantitated in the placental basal plate (BP) and villous trophoblast (TR), and compared to their expression in placenta accreta (n = 6). The proteins were also immunolocalized and quantitated in the accreta tissues. The messenger RNAs (mRNAs) of matrix metalloproteinase 9, -2, and tissue inhibitors of matrix metalloproteinase (TIMP)-1 were also measured. Results demonstrated that the BP and TR expressed low levels of RLN/RXFP1 and INSL4 through gestation. In accreta, increased RLN gene and protein in BP were associated with antepartum bleeding whereas INSL4 expression decreased throughout the TR. There were no changes in mRNAs for MMPs, but TIMP-1 was increased only in the invasive TR.

Identification of relaxin-responsive cells in the human choriodecidua at term.

The decidua of the human maternal-fetal interface is a local source of intrauterine relaxin, and the choriodecidua expresses its receptor (LGR7). Since these tissues consist of a variety of cells, we sought to identify the primary cell(s) responsible for LGR7 expression and relaxin responsiveness.

Relaxin and the human fetal membranes.

The human fetal membranes are complex tissues that perform many important functions during gestation. The extracellular matrix provides their strength to withstand the forces directed from the fetus and myometrium. Relaxin is a collagenolytic hormone that causes increased production of the matrix metalloproteinases. Its expression from the decidua is increased in patients with preterm premature rupture of the membranes, and its leucine-rich G receptor 7 is upregulated at preterm. The authors previously showed that relaxin is not involved in the infection-mediated cytokine response, but in the absence of infection, it causes increased secretion of both interleukin -6 and interleukin-8 from the membranes. In this article, the authors propose that relaxin is one of a number of sterile stimuli capable of causing increased proinflammatory cytokines, similar to but less robust than the effects of infection. These probably represent distinct inflammatory pathways involving different intracellular signaling events, which can result in either preterm premature rupture of the membranes or preterm labor. The current challenge is to fully understand these pathways and to clarify their similarities and differences.

Early placental insulin-like protein (INSL4 or EPIL) in placental and fetal membrane growth.

Early placental insulin-like protein (INSL4 or EPIL) is a member of the insulin superfamily of hormones, which is highly expressed in the placenta. We have confirmed this at term and shown it to be expressed by the maternal decidua. Although an abundance of locally acting growth factors are produced within the uterus during pregnancy, we hypothesized that INSL4 plays an important role in fetal and placental growth. We have demonstrated with cell lines and primary cells that it has a growth-inhibitory effect by causing apoptosis and loss of cell viability. We used primary amniotic epithelial cells for flow cytometry to show that INSL4 caused apoptosis, which was dose-related and significant (P < 0.05) at 50 ng/ml. This was confirmed by measurement of the nuclear matrix protein in the media. In comparison, relaxin treatment (up to 200 ng/ml) had no effect on apoptosis. The addition of INSL4 (3-30 ng/ml) also caused a loss of cell viability, although it had no effect on the numbers of cells at different phases of the cell cycle. Placental apoptosis is an important process in both normal placental development and in fetal growth restriction. Therefore, an in vivo clinical correlate was sought in fraternal twins exhibiting discordant growth. Expression of the INSL4 gene was doubled in the placenta of the growth-restricted twin compared to the normally grown sibling, suggesting that it may be linked to a higher level of apoptosis and loss of cell viability and, therefore, that it may contribute to fetal growth restriction.

Human decidual relaxin and preterm birth.

Relaxin in human pregnancy is both a systemic hormone from the corpus luteum and an autocrine/paracrine hormone at the maternal-fetal interface formed by the decidua/placenta and fetal membranes. We have focused our studies on the autocrine/paracrine roles of relaxin, especially in the preterm premature rupture of the fetal membranes, which causes 30-40% of preterm births. By using different techniques and different tissue collections, our laboratory has shown that expression of the relaxin genes and proteins in the decidua and placenta is increased in patients with preterm premature rupture of the fetal membranes. Relaxin binding and the expression of LGR7 are primarily in the chorion and decidua and are downregulated after spontaneous labor and delivery both at term and preterm. However, expression of LGR7 in the fetal membranes is significantly greater in all clinical situations at preterm than term, suggesting an important role for relaxin in these tissues at that time. The roles of the relaxin system in three potential causes of preterm birth are discussed: in the growth and proliferation of the membranes important for fetal membrane accommodation to fetal and placental growth, in acute infection, and in the inflammatory response leading to the initiation of labor.

Rendimentos dos cortes e não-componentes das carcaças de cordeiros terminados com dietas contendo diferentes fontes de óleo vegetal / Dressing of cuts and non-carcass components of lambs fed diets containing different sources of vegetal oil

Objetivou-se avaliar os rendimentos dos cortes e dos não-componentes das carcaças de cordeiros Santa Inês puros e ½ Dorset ½ Santa Inês, alimentados com dietas contendo diferentes fontes de óleo vegetal (óleo de soja, óleo de canola e óleo de linhaça) e uma dieta controle (sem adição de óleo vegetal). Após o abate, foram coletados sangue, pele, aparelho gastrintestinal cheio (esôfago + estômagos + intestinos delgado e grosso com seus conteúdos), aparelho gastrintestinal vazio (esôfago + estômagos + intestinos delgado e grosso, previamente esvaziados e limpos), aparelho reprodutor + bexiga, baço, fígado, coração, aparelho respiratório, rins com gordura perirrenal, cabeça, patas e cauda, que foram pesados para determinação do rendimento em relação ao peso vivo ao abate. Após resfriamento por 24 horas em câmara fria, pesou-se a carcaça e, posteriormente, dividiu-se longitudinalmente, sendo a metade esquerda seccionada em sete regiões anatômicas: perna, lombo, paleta, costelas flutuantes, costelas verdadeiras, baixos e pescoço. O estudo dos não-componentes da carcaça destacou a representabilidade dos pesos da pele (8,74 por cento) e do conteúdo gastrintestinal (10,65 por cento) na determinação do rendimento. As porcentagens dos cortes não apresentaram diferenças (p>0,05) em relação às dietas e grupos genéticos estudados.

Regulation of the human relaxin genes H1 and H2 by steroid hormones.

Relaxin, a peptide hormone important to the outcome of human pregnancy is expressed in a tissue specific manner as two genes known as relaxins H1 and H2, in addition to a third human relaxin H3, expressed primarily in the brain. The H1 and H2 genes are highly homologous, differentially expressed in reproductive tissues and appear to activate the same receptor, but their regulation is poorly understood. Based upon the known physiology of these hormones and the response elements in their 5'- and 3'-flanking regions, the possibility that progesterone and/or the glucocorticoids might influence their differential expression was therefore investigated. The changes in the mRNA levels of the relaxin genes in response to either medroxyprogesterone acetate (MPA) or dexamethasone (Dex) were analyzed by RT-PCR using a choriocarcinoma cell line (JAR) as a model system, because the expression of these genes in any primary human cell type is too low for such a study. The addition of 0.5 microM MPA to JAR cells, significantly upregulated the mRNA of only the relaxin H2, while the addition of 0.5 microM Dex significantly upregulated the mRNAs for both the relaxins, after 6h of treatment. Promoter assays indicated an early activation of transcription (1 h), which by 6 h had decreased. Progesterone and/or glucocorticoids could exert their effects via the GRE motif found on the 5'-flanking region of the relaxin genes. The H1-GRE differs from the H2-GRE by a single nucleotide, which may affect H1-GRE binding to the progesterone receptor (PR) but not the glucocorticoid receptor (GR). The antiprogestin RU486 inhibited the binding of the GR to both H1-GRE and H2-GRE, while it enhanced the binding of the PR to these GREs. As determined by gel shift assays, this GRE motif could bind to both the PR and GR and was therefore considered to be functional. Thus, both progesterone and glucocorticoids are capable of differentially regulating the expression of the two human relaxin genes in a model system.

Características de carcaça de cordeiros terminados em confinamento, com dietas contendo diferentes níveis protéicos / Carcass characteristics of lambs finished in drylot with diets contend different protein levels

Trinta e dois cordeiros (¹/2 Texel + » Bergamácia + » Corriedale), 16 machos inteiros e 16 fêmeas, foram alimentados durante 71 dias com dietas isoenergéticas (72 por cento NDT), variando os níveis protéicos (12, 16, 20 e 24 por cento PB) em baias individuais, com avaliaçäo de carcaça após abate. O rendimento verdadeiro médio da carcaça foi 54 por cento, enquanto o rendimento comercial médio foi 48 por cento. Verificou-se que os níveis protéicos entre 12 e 24 por cento näo afetaram o peso da carcaça quente e fria; o rendimento comercial da carcaça, o índice de compacidade da carcaça e do pernil, bem como a condiçäo corporal, conformaçäo, cobertura de gordura, consistência da gordura, cor da carne, espessura de gordura e deposiçäo de gordura. Os animais terminados com dietas com 12 por cento de proteína bruta foram os que apresentaram maior rendimento verdadeiro de carcaça

Relaxin causes proliferation of human amniotic epithelium by stimulation of insulin-like growth factor-II.

OBJECTIVE: The study was conducted to determine whether relaxin has a proliferative effect on amniotic epithelial cells and to show that this effect is caused by its stimulation of the insulin-like growth factor-II (IGF-II) gene. STUDY DESIGN: Immunolocalization and Northern analysis were used to confirm the expression of IGF-II by the fetal cells in the membranes. Human amniotic epithelial (WISH) cells were treated with doses of IGF-II or human relaxin and their proliferative effects measured. The mechanism of the effect of relaxin on cellular proliferation was studied with the use of an IGF-II-blocking antibody and Northern analysis for IGF-II gene expression after treatment with relaxin. An in vivo correlate was sought by quantitation of relaxin gene expression in 10 fetal membranes from women with normally grown and large for gestational age infants. RESULTS: The amniotic epithelial and cytotrophoblast cells of the fetal membranes expressed IGF-II, as did the amniotic epithelial-like (WISH) cell line. Treatment of WISH cells with IGF-II or relaxin caused a significant (P <.03) and dose-related increase in WISH cell proliferation over 5 days. The concurrent treatment with a blocking antibody to IGF-II significantly decreased the proliferative response to IGF-II (P <.002) and relaxin (P <.002). Treatment with relaxin caused a significant increase (P <.003) in the transcription of IGF-II in 24 hours. In fetal membranes, the levels of relaxin gene expression correlated with fetal membrane surface area (r = 0.76) and was significantly greater (P <.008) in the membranes from macrosomic infants (4020-4729 g) compared with those normally grown (2855-3830 g). CONCLUSION: IGF-II and relaxin both caused the proliferation of WISH cells. Concurrent treatment with an IGF-II-blocking antibody abrogated the proliferative effects of both hormones. Relaxin increased the transcription of IGF-II, and its expression levels in the fetal membranes correlated with the membrane surface area as well as neonatal birth weight. These data suggest that relaxin is a growth factor for the fetal membranes.

Decidual relaxins: gene and protein up-regulation in preterm premature rupture of the membranes by complementary DNA arrays and quantitative immunocytochemistry.

OBJECTIVE: This study was undertaken to show both decidual relaxin gene and protein up-regulation in preterm premature rupture of the fetal membranes. STUDY DESIGN: Membranes after preterm premature rupture (n = 4) have been matched in pairs with preterm intact membranes (n = 4). These tissues were from patients without infection, labor, preeclampsia or intrauterine growth restriction, and none of the patients had a latency period of more than 8 hours. The messenger RNAs from these tissues were used on complementary DNA expression arrays; 488 genes were analyzed. Relaxin gene expression was quantitated from the arrays and in additional tissues by Northern analysis. The two relaxin proteins, H1 and H2, in the decidual cells were immunolocalized and quantitated by microdensitometry with the use of specific antisera that were raised to decapeptides over the region of least homologic features. The expression of five other genes that were selected from the arrays were quantitated by Northern analysis. RESULTS: Relaxin gene expression was up-regulated 3.4-fold on the complementary DNA arrays but was not confirmed on Northern analysis. On the other hand, protein analysis for relaxin H1 and H2 in the decidual cells showed them to be significantly up-regulated (P <.0001, for both proteins) in patients with preterm premature rupture of the membranes compared with control subjects. The 20 most highly expressed genes at preterm in tissues without rupture were determined. In addition, analysis of the genes that were up-regulated with preterm rupture of the membranes showed 30 differentially expressed genes. CONCLUSION: Relaxin gene expression in the decidua is up-regulated, and its protein expression is significantly increased with preterm rupture of the fetal membranes.