Influence of different warming temperatures on the vitrified testicular fragments from pre-pubertal cat

Testicular vitrification is an alternative to preserve the genetic material of pre-pubertal animals. However, there are few studies on post-vitrification warming. Hence, the aim was to compare the influence of different warming temperatures on vitrified testicular fragments from pre-pubertal cats. The testicles were fragmented and divided into a control group (non-vitrified) and vitrified, using an association between dimethylsulphoxide and glycerol. The vitrified fragments were warmed at 50, 55 and 60°C/5 s. Morphological and morphometric evaluations were carried out using classical histology. Afterwards, the mitochondrial activity was evaluated using Rhodamine 123. The data were expressed in mean and standard error. The differences were considered significant when p < .05. In the histomorphological analysis, the testicular fragment presented seminiferous tubules with poorly developed germinal epithelium, compatible with pre-pubertal animals. The group warmed at 50°C presented similar to the control regarding the maintenance of the integrity of the tubules and cells, without stromal rupture and lamina propria alteration, as well as regarding the maintenance of the junctions between the cells. The group warmed at 55°C showed reduction of the cell junctions, and the one warmed at 60°C had increased detachment of the basement membrane (p < .05). The warming caused a reduction in the tubular diameter inversely proportional and progressive to the increase in temperature, with the highest diameter in the control group and the lowest in the 60°C group. The control group showed a lower incidence of Rhodamine 123, followed in ascending order of the warmings at 55 and 60°C. The higher mitochondrial activity was obtained with 50°C, showing an increase of the metabolic cell function at this temperature. It was concluded that the testicular fragment of pre-pubertal cats presents a better preserved morphology, morphometry and viability when warmed at 50°C.     

Supplementing organic-complexed or inorganic Co,Cu, Mn,and Zn to beef cows during gestation: physiological and productive response of cows and their offspring until weaning

One hundred and ninety non-lactating, pregnant beef cows (three-fourth Bos taurus and one-fourth Bos indicus; 138 multiparous and 52 primiparous) were assigned to this experiment at 117 ± 2.2 d of gestation (day 0). Cows were ranked by parity, pregnancy type (artificial insemination = 102 and natural service = 88), body weight (BW), and body condition score (BCS) and assigned to receive a supplement containing: 1) sulfate sources of Cu, Co, Mn, and Zn (INR; n = 95) or 2) an organic-complexed source of Cu, Mn, Co, and Zn (AAC; Availa 4; Zinpro Corporation, Eden Prairie, MN; n = 95). The INR and AAC provided the same daily amount of Cu, Co, Mn, and Zn, based on 7 g of the AAC source. From day 0 to calving, cows were maintained in a single pasture and were segregated three times weekly into 1 of the 24 individual feeding pens to receive treatments. Cow BW and BCS were recorded on days −30, 97, upon calving, and at weaning (day 367). Milk production was estimated at 42 ± 0.5 d postpartum via weigh–suckle–weigh (WSW) method. Liver biopsies were performed in 30 cows per treatment on days −30, 97, upon calving, and the day after WSW. Calf BW was recorded at birth and weaning. Liver and longissimus muscle (LM) biopsies were performed in 30 calves per treatment upon calving and 24 h later, the day after WSW, and at weaning. No treatment effects were detected (P ≥ 0.49) for cow BCS during gestation, despite AAC cows having greater (P = 0.04) BW on day 97. Liver Co concentrations were greater (P < 0.01) for AAC compared with INR cows, and liver concentrations of Cu were greater (P = 0.02) for INR compared with AAC cows on day 97. Upon calving, INR cows had greater (P ≤ 0.01) liver Cu and Zn concentrations compared with AAC cows. No other treatment differences were noted (P ≥ 0.17) for cow and calf liver trace mineral concentrations. Cows receiving AAC had greater (P = 0.04) hepatic mRNA expression of metallothionein 1A at calving, and their calves had greater (P = 0.04) hepatic mRNA expression of superoxide dismutase at weaning. Milk production did not differ between AAC and INR cows (P = 0.70). No treatment effects were detected (P ≥ 0.29) for mRNA expression of LM genes associated with adipogenic or muscle development activities in calves at birth and weaning. Calf birth and weaning BW also did not differ (P ≥ 0.19) between treatments. In summary, supplementing AAC or INR to beef cows during the last 5 mo of gestation yielded similar cow–calf productive responses until weaning.     

The effect of habitat fragmentation on communities of mutualists: Amazonian ants and their host plants

The consequences of fragmentation for communities of mutualist partners are for the most part unknown; moreover, most studies addressing this issue have been conducted on plant-pollinator communities. We evaluated how the experimental fragmentation of lowland Amazonian rain forest influenced a community of ant-plant mutualists. We inventoried a total of 1057 myrmecophytes in four fragments and four continuous forest sites; the twelve plant species recorded were occupied by 33 ant morphospecies, of which 11 were obligate plant inhabitants. Neither plant species richness, ant species richness, nor total ant-plant density were significantly lower in forest fragments. However, eight of the plant species, including three of the four most common, had higher mean densities in continuous forest than fragments. Of these four species, only one (Cordia nodosa) had significantly different colonization rates between habitat types, with higher colonization rates of plants in fragments. This may be because the Azteca species it is associated with increases in abundance in forest isolates. Although our results suggest that communities of ant-plant mutualists are likely to persist in fragmented tropical landscapes 25 years after fragment isolation, most species are rare and populations sizes in fragments are extremely low. Environmental and demographic stochasticity could therefore limit long-term population viability. We suggest future studies focus on evaluating how fragmentation has altered herbivore pressure and the dispersal of ants and plants to fragments, since the interaction of these factors is likely to have the greatest impact on long-term patterns of population persistence.     

Anti-Mullerian hormone and its relationship to ovulation response and fertility in timed AI Bos indicus heifers

This study evaluated the association between plasma anti-Mullerian hormone (AMH) concentration and fertility in Nelore (Bos indicus) heifers submitted to timed artificial insemination (TAI). At the onset of the synchronization protocol, heifers (n = 289) received a subcutaneous P4 ear implant (3 mg) and 2 mg of oestradiol benzoate. Eight days later, the P4 implant was removed and 0.5 mg of oestradiol cypionate, prostaglandin (0.265 mg, i.m.) and equine chorionic gonadotropin (300 UI, i.m.) was administered, and TAI was performed 48 hr after ear implant removal. Ovarian ultrasound evaluations were performed to measure number of ovarian follicles, dominant follicle size and ovulation response. Pregnancy diagnosis was performed by ultrasound 30 days after AI. Heifers with greater circulating AMH had more antral follicles, a smaller dominant follicle near timed ovulation and lower ovulation response to the timed AI protocol compared to heifers with lower circulating AMH. Although AMH and pregnancy outcome had a quadratic-shaped pattern, AMH was not significantly associated with fertility. In conclusion, heifers with lower AMH had larger follicles towards the end of the synchronization protocol and greater ovulation responses, whereas greater circulating AMH was unrelated to conception success.     

Effects of bovine leukemia virus infection on milk neutrophil function and the milk lymphocyte profile

The effects of bovine leukemia virus (BLV) on the immune response have been extensively investigated; however, its effects on mammary gland immunity are only speculative. Although BLV has a tropism for B cells, it can affect both adaptive and innate immunities because these systems share many effector mechanisms. This scenario is the basis of this investigation of the effects of BLV on mammary gland immunity, which is largely dependent upon neutrophilic functions. Thus, the present study sought to examine neutrophilic functions and the lymphocyte profile in the milk of naturally BLV-infected cows. The viability of the milk neutrophils and the percentage of milk neutrophils that produced reactive oxygen species (ROS) or phagocytosed Staphylococcus aureus were similar between BLV-infected and BLV-uninfected dairy cows. Furthermore, the expression of CD62L and CD11b by the milk neutrophils and the percentage of milk neutrophils (CH138⁺ cells) that were obtained from the udder quarters of the BLV-infected cows were not altered. Conversely, the median fluorescence intensity (MFI) representing intracellular ROS production and the phagocytosis of S. aureus, the expression of CD44 by the milk neutrophils and the percentage of apoptotic B cells were lower in the milk cells from BLV-infected dairy cows, particularly those from animals with persistent lymphocytosis (PL). The lymphocyte subsets were not different among the groups, with the exception of the percentage of CD5⁻/CD11b⁻ B cells, which was higher in the milk cells from BLV-infected cows, particularly those with PL. Thus, the present study provides novel insight into the implications of BLV infection for mammary gland immunity.     

Management strategies and distribution of Aphanomyces root rot of alfalfa (Medicago sativa), a continuing threat to forage production in the United States

Alfalfa (Medicago sativa) is one of several legumes that is affected by Aphanomyces root rot (ARR) caused by Aphanomyces euteiches. Symptoms of ARR on alfalfa seedlings include a yellow-grey discolouration of roots, rotting and loss of lateral roots, stunted growth, chlorotic foliage and reduction of nitrogen-producing nodules on roots. Infection can also occur on adult plants leading to loss of lateral roots and nodules. At the seedling stage, ARR decreases alfalfa stand establishment, and field longevity is reduced when adult plants are infected. A. euteiches is an oomycete pathogen that has motile zoospores and thick-walled oospores that can survive for many years in soil. Two races are currently recognized by pathogenicity on differential alfalfa check cultivars. Most alfalfa cultivars contain race 1 resistance, but there is an increasing development of cultivars with resistance to race 2. Management strategies include planting resistant cultivars, avoiding planting in fields with poor drainage and rotating crops with nonhost plants.     

Potential Utilization of Green Tide-Forming Macroalgae from Patos Lagoon,Rio Grande-RS,Brazil

ABSTRACT

This study was conducted to evaluate the biochemical and nutritional characteristics of naturally occurring estuarine macroalgae in the Patos Lagoon (Rio Grande, Brazil). The study of these macroalgae, dominated by Ulva and Cladophora species, contributes to current research on the potential utilization of green tide-forming algae. Although high levels of carotenoids (378 ± 0.02 µg g^?1) and antioxidant activity (75.0% for DPPH) were found in the naturally occurring macroalgae, these macroalgae cannot be used in the food industry due to the presence of toxic levels of arsenic (exceeding the thresholds of both French and Brazilian legislation for food). Regarding the potential of the studied macroalgae as a biofuel source, the low lipid and very high carbohydrate contents suggest a possible use in the fermentation of ethanol, butanol, or methane.     

Estimating bergamot juice adulteration of lemon juice by high-performance liquid chromatography (HPLC) analysis of flavanone glycosides

The chemical composition of 30 samples of juices obtained from bergamot (Citrus bergamia Risso and Poit.) fruits is reported and compared to the genuineness parameters adopted by Association of the Industry of Juice and Nectars (AIJN) for lemon juice. It was found that the compositional differences between the two juices are distinguishable, although with difficulty. However, these differences are not strong enough to detect the fraudulent addition of bergamot juice to lemon juice. Instead, we found the high-performance liquid chromatography (HPLC) analysis of the flavanones naringin, neohesperidin, and neoeriocitrin, which are present in bergamot juice and practically absent in the lemon juice, is a convenient way to detect and quantify the fraudulent addition of bergamot juice. The method has been validated by calculating the detection and quantification limits according to Eurachem procedures. Employing neoeriocitrin (detection limit = 0.7 mg/L) and naringin (detection limit = 1 mg/L) as markers, it is possible to detect the addition of bergamot juice to lemon juice at the 1% level. When using neohesperidin as a marker (detection limit = 1 mg/L), the minimal percentage of detectable addition of bergamot juice was about 2%. Finally, it is reported that the pattern of flavonoid content of the bergamot juice is similar to those of chinotto (Citrus myrtifolia Raf) and bitter orange (Citrus aurantium L.) juices and that it is possible to distinguish the three kinds of juices by HPLC analysis.     

Growth of Phlebiopsis gigantea in wood of seven conifer species

Heterobasidion parviporum and Heterobasidion annosum are widely distributed root‐rot fungi that infect conifers throughout Europe. Infection of conifer stumps by spores of these pathogens can be controlled by treating fresh stumps with a competing non‐pathogenic fungus, Phlebiopsis gigantea. In this study, growth of three Latvian strains of P. gigantea and the biological control agent ‘Rotstop’ strain was evaluated in stem pieces of Norway spruce, Scots pine, lodgepole pine, Douglas‐fir, Weymouth pine, Siberian larch and Sitka spruce. The growth rates of one H. parviporum and one H. annosum isolate were also measured in the same stem pieces. The growth rate of P. gigantea varied greatly in wood of different conifer species. It was higher in the three pine species, lower in Norway spruce and lowest in Sitka spruce and Siberian larch, and in Douglas‐fir, this fungus did not grow. The largest area of wood occupied by P. gigantea was in lodgepole pine. Growth of Latvian isolates of P. gigantea in the wood of Pinus and Picea species was comparable to that of the Rotstop isolate. Consequently, stump treatment with local P. gigantea isolates should be recommended. However, our results suggest that Douglas‐fir stump treatment against Heterobasidion by P. gigantea may be ineffective and other stump treatment methods should be considered.     

Pollination and in vitro germination of seeds for interspecific hybridization of <Emphasis Type="Italic">Psidium guajava</Emphasis> and <Emphasis Type="Italic">Psidium cattleianum</Emphasis>

The genus Psidium includes important fruit crops. However, there are very few studies focusing on its reproductive biology, which limits the establishment of breeding programs. The present work investigated the reproductive biology of Psidium guajava and Psidium cattleianum in terms of compatibility of crossings between these two species aiming at interspecific hybridization because the latter species is an important source of resistance against the nematode Meloidogyne enterolobii. Several types of crosses were performed to understand the reproductive biology of these species, including the compatibility of intra- and interspecific crossings, using assisted in vivo germination of pollen grains on the stigma. In addition, the in vitro germination of both Psidium species was studied at different stages of fruit development to rescue young seeds to improve the chances of obtaining the hybrids. No fruits of 270 pollinations were obtained on guava buds at the pre-anthesis stage, regardless of the source of the pollen grain and the cultivar used as female genotype. Microscopic analyzes demonstrated the germination of pollen grains and pollen tube growth at crosses between guava cv. ‘Pedro Sato’ (P. guajava) and Psidium cattleianum. High germination percentages of Psidium cattleianum seeds were obtained in MS medium without sucrose or containing 15 g/L of this carbohydrate.