Sea turtle strandings reveal high anthropogenic mortality in Italian waters

• 1. Spatio‐temporal distribution and anthropogenic mortality factors were investigated in loggerhead turtles (Caretta caretta) found stranded or floating in the waters around Italy. A total of 5938 records for the period 1980–2008 were analysed concerning loggerhead turtles measuring from 3.8 to 97 cm curved carapace length (mean: 48.3 cm).
• 2. Results highlighted the following conservation issues: (i) in the study area, anthropogenic mortality is higher than natural mortality; (ii) interaction with fisheries is by far the most important anthropogenic mortality factor; (iii) longlines are an important mortality factor in the southern areas; (iv) trawlers are the cause of high numbers of dead strandings in the north Adriatic; (v) entanglement in ghost‐gear or in other anthropogenic debris affects high numbers of turtles; and (vi) boat strikes are an important source of mortality in most areas but mostly in the warm seasons.
• 3. Results also indicate that: (vii) the north Adriatic is the area with the highest turtle density; and (viii) the south Adriatic and to a lesser extent the surrounding areas of the north Adriatic and the Ionian, are important developmental areas for loggerhead turtles in the first years of life.
• 4. Italy is in the centre of the Mediterranean Sea and borders major foraging areas for the loggerhead turtles in the region, and these results confirm previous concerns about the level of anthropogenic mortality in Italian waters. Copyright © 2010 John Wiley & Sons, Ltd.

     

Portable surface plasmon resonance immunosensor for the detection of fluoroquinolone antibiotic residues in milk

An inexpensive and portable surface plasmon resonance (SPR) sensor, SPReeta Evaluation Kit SPR3, has been used to develop a biosensor for the determination of fluoroquinolone antibiotics (FQs) and to demonstrate its performance analyzing FQ residues in milk samples. The SPReeta three-channel gold chips were activated with a mixed self-assembled monolayer (m-SAM) and functionalized with a FQ haptenized protein. Binding of the antibody produced a concentration-dependent increase of the SPR signal as a result of the change in the refraction index. Similarly, the presence of the FQ produced a dose-dependent decrease of the response, which allowed a good limit of detection (LOD) to be obtained (1.0 ± 0.4 μg L(-1) for enrofloxacin in buffer). The response was reproducible in all three channels, on different injections and days, and also between chips. Milk samples could be analyzed after a simple sample treatment involving fat removal by centrifugation and dilution with water. Under these conditions calibration curves were obtained showing that FQ residues can be analyzed in milk samples with an IC(50) value of 26.4 ± 7.2 μg L(-1) and a LOD of 2.0 ± 0.2 μg L(-1) (for enrofloxacin), far below the European Union regulations for this antibiotic family in this matrix. Finally, the paper also demonstrates that the biosensor is able to selectively detect the presence of FQs in milk samples, even in the presence of other antibiotics. Enrofloxacin, ciprofloxacin, and norfloxacin residues were detected in blind samples supplied by Nestle? Co.     

Comparison of different media for the isolation of small ruminant strains of Mycobacterium paratuberculosis

Two different egg based media, with and without the incorporation of sodium pyruvate, were used to isolate M. paratuberculosis from sheep, goat and cattle samples obtained at our laboratory for two years. Both media were adequate for bovine material, with a slightly improved isolation rate for Herrold's egg yolk medium incorporating sodium pyruvate; however, most of the small ruminant strains grew only on L?wenstein-Jensen medium without sodium pyruvate. Those results point out the need to use different media when working with small ruminant samples and provide further evidence for the existence of different varieties of M. paratuberculosis causing paratuberculosis in cattle and in small ruminants.     

PULSED WAVE-DOPPLER ULTRASONOGRAPHIC EVALUATION OF THE COMMON CAROTID ARTERY IN THE RESTING HORSE: PHYSIOLOGIC DATA

A pulsed wave-Doppler ultrasonographic evaluation of common carotid arterial blood flow was carried out on 63 healthy Italian Saddlebred horses. Vessel diameter and tracing morphology were evaluated were evaluated and blood flow parameters (systolic, diastolic and mean velocity, acceleration and deceleration of the systolic wave, cartid pulse volume) were calculated and correlated with class variables (sex, age and body weight). On the basis of the presence of an incisure in the ascending branch of the systolic curve, subjects were divided in two groups: one with a bifid systolic curve and the other with a monophasic aspect. Correlations between: 1) diameter of the vessel and body weight and 2) carotid pulse volume and flow velocity (systolic, diastolic and mean) were found. A greater systolic pulse volume was found in male subjects, in subjects with greater body weight and in those which had a monophasic systolic wave.     

Relative contributions of hyperplasia and hypertrophy to growth in cattle

The relationship between postweaning DNA accumulation and body weight was calculated for a reference growing steer. Accumulation of DNA (hyperplasia) was calculated from patterns of protein accretion derived from literature data and protein-to-DNA ratios (Pro/DNA) measured in samples of muscles from chuck, round and plate, hides, intestines, rumens and livers of steers slaughtered at body weights ranging from 150 to 700 kg. Amounts of protein relative to DNA found in muscles from chuck, round and plate were used to estimate Pro/DNA changes in carcass. Corresponding values from intestines, rumen and liver were used to estimate Pro/DNA patterns for viscera. This ratio increased in carcass until animals reached body weights of approximately 300 kg and leveled off thereafter. No cell enlargement was evident in viscera. Thus, postweaning protein accretion in carcass results from both cell enlargement (hypertrophy) and DNA accumulation (hyperplasia) until body weights of 300 kg are attained, subsequent growth appeared to be hyperplastic in nature. In viscera, hyperplasia was the primary determinant of protein accretion.     

Genetic and antigenic characterization of the surface lipoprotein P48 of Mycoplasma bovis

The presence of a membrane lipoprotein homologous to the P48 of Mycoplasma agalactiae was investigated in different Mycoplasma bovis isolates selected by geographical locations and biological properties. Its potential as a diagnostic tool was also discussed. The presence of a specific signal observed in all M. bovis field isolates probed with a rabbit antiserum raised against the M. agalactiae recombinant P48 demonstrated that this protein is structurally and antigenically conserved within the M. bovis cluster. No signal was detected when testing six different mycoplasma species found in cattle. The p48 gene was identified by PCR approach and partially sequenced. Full length gene sequence was obtained by direct bacterial chromosome sequencing. Five UGAs were selectively mutated into UGG and the full length mutated gene, lacking the signal peptide, was cloned and expressed in Escherichia coli. The purified recombinant antigen (r-P48) was evaluated as a potential marker of infection using a panel of 86 well-characterized sera from experimentally and naturally infected cattle. Specific IgM antibodies were detected within 6-9 days after experimental infection followed by an IgG response lasting from the third/fourth week after contact. Although antibody titers were well below those observed in sheep or goats infected with M. agalactiae, results suggest that M. bovis r-P48 can be used as a specific marker of infection.     

Primary hyperoxaluria (L-glyceric aciduria) in a cat

A 7-month-old, male European cat was examined because of weakness and inappetence. The cat was dehydrated, polypnoeic and severely weak. Severe, generalised muscle atrophy was present. Spinal reflexes were all decreased to absent. Blood analysis and urinalysis showed several abnormalities, including intermittent hyperoxaluria. The L-gliceric acid concentration was remarkably increased. Electrodiagnostic tests of the peripheral nervous system were abnormal. At necropsy, generalised muscle atrophy was observed. Microscopically, both kidneys showed intraluminal birefringent oxalate crystals. Motor neuron degeneration and accumulation of neurofilaments were observed in the axons of the spinal motor neurons.     

High-frequency ultrasonography of the skin of clinically normal dogs

OBJECTIVE: To assess the applicability of high-frequency diagnostic ultrasonography for evaluation and accurate measurement of the skin thickness of clinically normal dogs. ANIMALS: 26 healthy dogs (12 sexually intact males, 13 sexually intact females, and 1 spayed female) of various breeds and ages. PROCEDURE: Ultrasonographic examination of the skin and histomorphometric analysis of skin biopsy specimens obtained from the same site were performed. A 13-MHz linear-array transducer was used to obtain a series of ultrasonographic images of the skin in the flank region; images were analyzed and measured by use of imaging software. Cutaneous biopsy specimens were placed in fixative and then stained with H&E and Masson trichrome stains. Histomorphometric analysis was performed by use of an image analyzer. Thickness of the epidermis and dermis of each specimen was evaluated by use of a semiautomatic procedure of quantification. Data obtained from ultrasonographic and histologic measurements were compared by use of the Pearson correlation test. RESULTS: The ultrasonographic pattern of canine skin was consistently characterized by 3 distinct, defined echogenic layers corresponding to the epidermal entry echo, epidermis and dermis, and subcutaneous tissues. A positive correlation was found between ultrasonographic and histologic measurements of skin thickness. CONCLUSIONS AND CLINICAL RELEVANCE: Comparison between ultrasonographic and histologic appearance of the skin revealed that layering of canine skin (ie, epidermis and dermis) and the subcutaneous tissues may be recognized and measured by use of high-frequency ultrasonography. Thus, diagnostic ultrasonography may be a useful tool for the noninvasive evaluation of cutaneous disorders in dogs.     

Immunohistochemical detection of growth hormone (GH) in canine hepatoid gland tumors

The aim of this study was to detect immunohistochemically means growth hormone (GH) in 24 hepatoid gland adenomas and 5 hepatoid gland carcinomas and to compare the difference of immunoreactivity between types of tumors. The tumors were classified according to the WHO standards. Tissue sections which were prepared from formalin-fixed, paraffin wax-embedded tissues from 25 male and 4 female dogs were carried out immunostaining using polyclonal primary anti-hGH and EnVision method. Of 24 hepatoid gland adenomas (perianal gland adenomas) 23 (95.8%) were positive. All 5 hepatoid gland carcinomas (perianal gland carcinomas) were positive. No statistically significant differences in percentage of labelled cells between malignant and benign tumors were seen. The present demonstration of GH in hepatoid gland tumors adds new data on GH in extra-pituitary tissues and hormon-dependent tumors.