Chemical Composition and Sulfur Speciation in Bulk Tissue by X-Ray Spectroscopy and X-Ray Microscopy: Corneal Development during Embryogenesis

The chemical composition and sulfur (S) speciation of developing chick corneas at embryonic days 12, 14, and 16 were investigated using synchrotron scanning x-ray fluorescence microscopy and x-ray absorption near-edge structure spectroscopy. The aim was to develop techniques for the analysis of bulk tissue and identify critical physicochemical variations that correlate with changes in corneal structure-function relationships. Derived data were subjected to principal component analysis and linear discriminant analysis, which highlighted differences in the elemental and S species composition at different stages of embryonic growth. Notably, distinct elemental compositions of chlorine, potassium, calcium, phosphorus, and S altered with development during the transition of the immature opaque cornea to a mature transparent tissue. S  structure spectroscopy revealed developmentally regulated alterations in thiols, organic monosulfides, ester sulfate, and inorganic sulfate species. The transient molecular structures and compositional changes reported here provide a deeper understanding of the underlying basis of corneal development during the acquisition of transparency. The experimental and analytical approach is new, to our knowledge, and has wide potential applicability in the life sciences.     

Mode of delivery is associated with different hematological profiles in the newborn calf

Several studies on babies have shown that the type of delivery can influence the hematological and immune status of the newborn. In bovine medicine, some authors reported the hematological pattern of the newborn calf, but never related it with the calving process or other perinatal factors. The purpose of the present study was to evaluate the hematological profile in newborn calves in relation to the type of delivery. A total of 41 healthy calves were enrolled; 16 Friesian calves which were born by vaginal delivery without assistance (VD), and 25 Belgian Blue calves that were born by elective Caesarean section (CS). As soon as the calves were born, a complete clinical examination was performed to verify viability and maturity. At 10 min after birth, 2 mL venous blood was collected to perform the blood gas and acid-base evaluation. Blood samples were subsequently collected from the jugular vein within 30 min after birth, and at 1, 2, 3, 7, and 14 days of age. An automatic analyzer was used to determine hemoglobin concentration (Hb), hematocrit (Ht), and red and white blood cell counts, while differential leukocyte count was performed microscopically. Statistical analysis was applied to assess differences between the groups and within the group for all parameters between each sampling time (P ≤ 0.05). All the calves were born alive, viable, and mature. There were no acidotic calves, but statistical analysis revealed many differences, as higher pH, base excess (BE) (P ≤ 0.05), PO₂ (P < 0.001), and sO₂ (P < 0.0001) in the VD group. Levels of hemoglobin concentration, hematocrit, and red blood cell number were constantly higher in CS calves (P < 0.001). In comparison with the VD calves, white blood cell and neutrophil absolute number were higher at birth and at 14 days of age in the CS group (P < 0.001 and P ≤ 0.05). The mode of delivery, therefore, seems to have an influence on the oxygenation levels and on the hematological and nonspecific immunity profile of the newborn calf.     

Luteal function in jennies following PGF(2alpha) treatment 3 days after ovulation

Native PGF(2alpha) and its analogs have been used in the horse mare to manipulate ovarian activity, primarily as luteolytic agents to induce estrus. Despite numerous studies on the effects of these luteolysins in the mare, to date only a single investigation has been conducted in the jenny. The aim of this study was to evaluate the response of the corpus luteum (CL) to a single dose of PGF(2alpha) given 3 days (72h) after ovulation and to establish the plasma progesterone (P4) profile from pre-treatment to post-treatment ovulation in the Martina Franca donkey. Twenty-two jennies were ultrasonographically monitored and treated 72h after the detection of ovulation with 0.075 mg i.m. of R-cloprostenol. From the day of ovulation until ovulation post-treatment, blood was collected daily for P4 determination by enhanced luminescence immunoassay. All the jennies except one, exhibited behavioral signs of PGF(2alpha)-induced estrus within 4 days of treatment lasting 5.4+/-1.16 days. Post-treatment ovulation was also hastened, reducing the interovulatory interval (9.6 days). In response to treatment, plasma P4 concentrations fell to estrus levels and then remained constant until the next ovulation in all but the non-responding animal. Our findings indicate that PGF(2alpha) treatment on Day 3 post-ovulation causes the functional regression of the CL in the jenny, reflected both by the rapid induction of estrus and ovulation and by an abrupt drop in circulating P4 concentrations.     

A rapid altitudinal range expansion in the pine processionary moth produced by the 2003 climatic anomaly

Climatic anomalies may produce, or accelerate, geographic range expansions of species limited by temperature or other climatic variables. Most such expansions are only temporary, before the prevailing climatic conditions drive the founder populations extinct. In contrast, here, we report a recent rapid shift of the range limit during the record hot summer of 2003 in southern Europe that has the potential to be both permanent, and to have important implications on species range dynamics in general. The winter pine processionary moth (Thaumetopoea pityocampa), an important pine defoliator whose larvae feed in colonies during the winter, is limited in its distribution by winter temperatures. In the last three decades, warmer winters have led to a gradual but substantial expansion of its range both latitudinally and altitudinally. In the summer of 2003, T. pityocampa underwent an extraordinary expansion to high elevation pine stands in the Italian Alps; its altitudinal range limit increased by one third of the total altitudinal expansion over the previous three decades. In an experiment, we found flight activity of newly emerged females to increase with temperature. By determining a threshold temperature for flight take‐offs under controlled conditions, we calculated that the nights above the threshold temperature were over five times more frequent, and considerably warmer, at the range limit in 2003 than in an average year. We therefore attribute the colonization of extreme, high‐elevation sites to increased nocturnal dispersal of females during the unusually warm night temperatures in June – August 2003. Importantly, the colonies established at extreme sites survived the winter and produced offspring in 2004, although the range did not expand further because of low night temperatures that year. We discuss several life‐history characteristics of T. pityocampa that maximize the likelihood of population persistence at the new range limit. As global warming continues and climatic anomalies are predicted to become more frequent, our results draw attention to the importance of extreme climatic events in the range formation of phytophagous insects.     

Trapping sand flies (Diptera: Psychodidae) in the Emilia-Romagna region of northern Italy.

The efficiency and practicality of two trapping methods for adult Phlebotomine sand flies in two areas of the Emilia-Romagna region (Italy) were evaluated. Suction traps (CO2) and sticky traps (ST) were used to collect sand flies every two weeks, from June to September, 1999, from 16:00 to 07:00. Two CO2 traps were activated at the same time for each area (one with light and one without light), whereas 38 (four with light and 34 without lights) and 48 (four with light and 44 without) sticky traps were activated in Borghi and Longiano, respectively. An Index of Apparent Abundance (IAA) was calculated for each trap type and area. A total of 2,253 sand flies was trapped over the four-month period, with 1,765 collected from Borghi and 488 from Longiano. Phlebotomus perfiliewi was the most abundant species collected, comprising 99.6% and 84.6% of the total flies trapped in Borghi and Longiano, respectively. Other species were also collected within the two areas (Phlebotomus perniciosus and Phlebotomus mascittii) but were not considered for further analyses due to low catches. Significantly more specimens were caught using CO2 than sticky traps and the addition of a light source also improved the catches, however, a significantly greater number of female specimens were collected by a CO2 trap without a light source. Phlebotomus perfiliewi thus appears to show a photophobic reaction in the case of females when trapped using CO2/light attractants.     

Bacterial citrate synthase expression and soil aluminum tolerance in transgenic alfalfa

Alfalfa is very sensitive to soil acidity and its yield and stand duration are compromised due to inhibited root growth and reduced nitrogen fixation caused by Al toxicity. Soil improvement by liming is expensive and only partially effective, and conventional plant breeding for Al tolerance has had limited success. Because tobacco and papaya plants overexpressing Pseudomonas aeruginosa citrate synthase (CS) have been reported to exhibit enhanced tolerance to Al, alfalfa was engineered by introducing the CS gene controlled by the Arabidopsis Act2 constitutive promoter or the tobacco RB7 root-specific promoter. Fifteen transgenic plants were assayed for exclusion of Al from the root tip, for internal citrate content, for growth in in vitro assays, or for shoot and root growth in either hydroponics or in soil assays. Overall, only the soil assays yielded consistent results. Based on the soil assays, two transgenic events were identified that were more aluminum-tolerant than the non-transgenic control, confirming that citrate synthase overexpression can be a useful tool to help achieve aluminum tolerance. Pierluigi Barone and Daniele Rosellini contributed equally to this work.     

Characteristics of cryptic/ectopic and contralateral scrotal testes in dogs between 1 and 2 years of age

Testicular malposition represents a common developmental genital defect in dogs and can affect one or both testes. In both humans and dogs, unilateral cryptorchism is more frequently detected and thought to be the expression of a genetic abnormality affecting both the undescended and scrotal testis. In the dog, there is evidence of degenerative processes affecting the maldescended testis. However, the histologic and functional changes that occur in the scrotal testis of unilateral cryptorchid or ectopic individuals remain a source of debate. Because the bilateral surgical removal of the testes leads to some undesirable side effects, the aim of this study was to evaluate the necessity for performing bilateral orchiectomy in young unilateral cryptorchid dogs. A morphologic study of both cryptic/ectopic and scrotal testes in young dogs affected by unilateral testicular maldescent was therefore conducted. The study was conducted on 10 dogs aged 1 to 2 yr and affected by unilateral testicular maldescent. We found that, in young dogs, even if no neoplastic lesions were observed, morphologic abnormalities are detectable between 1 and 2 yr of age in the maldescended testes with severity dependent on testicular position. In contrast, in the scrotal testes, the histologic and immunohistochemical exam failed to find signs of incorrect development or morphologic abnormalities. The results seem to suggest that, though the early removal of the undescended testis is recommended, continuous monitoring of the scrotal testis for the life of the dog is preferable to removing it considering the undesirable side effects related to castration.     

An Apgar scoring system for routine assessment of newborn puppy viability and short-term survival prognosis

The Apgar scoring system is an easy and reliable method for evaluating both human and animal neonates. However, its use is not widespread in veterinary medicine. The current study assessed a modified Apgar scoring system for routine evaluation of newborn puppies. Heart rate, respiratory effort, reflex irritability, motility, and mucus color have been evaluated in the score. Specifically, we used 5 min after birth Apgar score to assess newborn viability and short-term survival prognosis, as well as related characteristics, in 193 puppies from 42 litters, 65 born by spontaneous delivery, 66 by assisted delivery, and 62 by cesarean section. The percentage of puppies that were dead 2 h after birth was higher in the 4 to 6 Apgar score group versus that in the 7 to 10 score group (P < 0.01) and in the 0 to 3 score group versus that in the 7 to 10 score group (P < 0.0001). Delivery method did not affect survival. There was a marked reduction in the number of puppies searching for the mammary gland in the 0 to 3 and 4 to 6 Apgar score groups compared with that in the 7 to 10 score group (P < 0.0001); there was a difference between the 0 to 3 and the 4 to 6 score groups as well (P < 0.05). Suckling/swallowing reflexes were present in fewer puppies in the 0 to 3 and 4 to 6 score groups compared with that in the 7 to 10 group (P < 0.0001), with no significant differences between the 0 to 3 and the 4 to 6 score groups.