Helminth fauna in roe deer (Capreolus capreolus Linnaeus, 1758) in the province of Grosseto (central Italy).

Helminth infection was analysed at necropsy and coprology in a total of 54 roe deer from the province of Grosseto (central Italy) between 2018 - 2020. Age and sex data were recorded for each deer for a total of 31 adults (23 females, 8 males) and 23 juveniles (11 females, 12 males). The results on the small intestine (51 samples) highlighted that nematodes belonging to the species Trichostrongylus colubriformis were the most prevalent parasite (41.2 %), followed by the cestode Moniezia expansa (7.8 %). The large intestine results (52 samples) showed Trichuris spp. (53.8 %), Oesophagostomum venulosum (50 %) and Chabertia ovina (26.9 %). In the abomasum, only Ostertagia ostertagi (17.9 %) was found. Of the 34 samples analysed by bronchopulmonary, only the lung of an adult female was positive for Dictyocaulus spp. In two livers out of 33 samples analysed, nematodes of the species Setaria tundra were found on the surface. Copropositivity was observed in 45 of the 52 faecal samples analysed. The results of the present study indicate that the roe deer is host to several species of parasites, which are also common in other cervids and domestic ruminants. Statistical testing highlighted a significant difference between mean intensities in males and females.

Canine filarial infections in Liguria, north-west Italy.

This paper reports the findings of a study on the presence of various species of filarial nematodes in dogs in Liguria, north-west Italy, a region traditionally considered free from the disease. Between 2009 and 2012 blood samples were taken from 365 dogs in rural areas in Liguria. The blood samples were then submitted to Knott's test, histochemical staining, polymerase chain reaction (PCR) and the enzyme-linked immunosorbent assay (ELISA) for Dirofilaria immitis antigens. Overall, 35 of the 365 dogs were positive using Knott's test for microfilariae (prevalence 9.6%; 95% confidence interval (CI): 6.6-12.6%). Acanthocheilonema reconditum was the most prevalent species (8.0%), while Dirofilaria repens (1.4%) and Dirofilaria immitis (0.6%) were less common. One co-infection by D. repens and A. reconditum was observed. All morphological identifications were confirmed by histochemical staining and PCR. In addition, a retrospective analysis of data on D. immitis antigens in 11,363 samples of canine sera was carried out. Sera were collected and analysed for D. immitis antigens by the Istituto Zooprofilattico Sperimentale (IZS) of Piedmont, Liguria and Aosta Valley (Imperia section) between 2004 and 2013 during annual tests for leishmaniasis on autochthonous dogs throughout Liguria. Serological data from IZS showed an overall seroprevalence of 0.65% (95% CI: 0.50-0.80%) for D. immitis throughout the region. The present study updates the epidemiological map of canine filarial infections in Italy and suggests the need for surveillance and prophylaxis in Liguria.

Canine filarial infections in Tuscany, central Italy.

This study was conducted in order to investigate the epidemiology of filarial species in a region of central Italy (Tuscany) in dogs that did not undergo prophylaxis for filariasis. From 2007 to 2009, 630 blood samples were collected from 40 kennels throughout the regional territory. Samples were analysed with Knott's modified test and with an enzyme-linked immunosorbent assay (ELISA) for the detection of Dirofilaria immitis antigens, those positive for microfilariae were also subjected to histochemical staining for acid phosphatase activity to validate the identification of the species. An overall elevated prevalence of filariasis (23.2%) was found. Dirofilaria immitis and D. repens were found to be the dominant species, with similar prevalences (12.5% and 12.1% respectively), while Acanthocheilonema reconditum was less common (1.9%). Results of this study indicate that monitoring for filariasis should be kept at a high level. A constant monitoring of the canine sanitary status should also be carried out for the protection of human health, considering the zoonotic potential of filarial worms.

Effects of copper on larvae of the marbled crab Pachygrapsus marmoratus (Decapoda, Grapsidae): Toxicity test and biochemical marker responses.

The importance of trace elements in ecotoxicological investigations is a well-known issue when monitoring polluted areas such as commercial harbors. Copper represents one of the most common metal contaminants, often detected in these areas as it is widely employed in various fields and has many sources of inflow in the marine environment. Pachygrapsus marmoratus is a widespread intertidal crab species that has been extensively studied in ecology, ethology and population genetics. Ecotoxicological studies have also been performed, exclusively on the adult stage. In the present study we investigated the mortality and biochemical (oxidative stress and neurotoxicity) responses of P. marmoratus larvae exposure to environmental relevant concentration of copper. Results showed dose-dependent responses in terms of larval mortality, with a calculated LC50 value of 0.5 mg/L of Cu2+. The LC50 concentration was used as the starting point for subsequent biochemical response evaluation. Results also demonstrated dose-dependent activation of antioxidant systems assuming a compensatory antioxidant activity to prevent higher cellular damage when larvae were exposed to the highest concentrations of copper. Moreover, a significant enhancement of neurotransmitter activities was observed, assuming a possible direct interaction of copper with the enzymes or an increase of free copper ion aliquot into the cells.

Antenatal diagnosis of thalassemia major in Sardinia.

In this report, we summarized our experience, carried out in Sardinia, with antenatal diagnosis in one thousand pregnancies in which the fetus was at risk for homozygous beta-thalassemia. In the majority of these cases, the thalassemia lesion segregating in the family was the nonsense mutation at the codon corresponding to amino-acid 39. At the outset (976 cases) we used globin chain synthesis analysis by column chromatography on fetal blood obtained by placental aspiration, and recently (24 cases) we employed the synthetic oligonucleotide method on amniocyte DNA. Apart from 126 pregnancies still in progress, in all the other cases the diagnosis has been confirmed. In the majority of the cases (99%), we obtained sufficient fetal blood for the analysis. The fetal mortality associated with placental aspiration was 6.1%. The biochemical analysis gave reliable results. We had two misdiagnoses (0.2%): one due to a nonglobin protein comigrating with the beta chains and the other for a misclassification of the type of thalassemia segregating in the family. The oligonucleotide method gave clear-cut results in all the cases tested. The method was sensitive enough to detect the mutation directly in the DNA isolated from 20-25 ml of amniotic fluid in 75% of the pregnancies tested. In one case, we successfully employed this method for the analysis of the DNA isolated from chorionic villi. The oligonucleotide method seems to be the best procedure for monitoring the pregnancies at risk for beta-thalassemia in places where one or a few beta-thalassemia lesions are prevalent.

Antenatal diagnosis of beta-thalassemia in Sardinia.

This paper reviews the characteristics and the results of 15 years of experience with a preventive program, based on carrier screening and prenatal diagnosis, designed to control thalassemia major in the Sardinian population. The education of the population about thalassemia and the modalities for its prevention was accomplished via the mass media. Carrier screening was carried out voluntarily on couples of child-bearing age. Prenatal diagnosis was initially carried out by fetal blood analysis; since 1983, it has been done by DNA analysis on non-amplified or amplified DNA. Different chorionic villous sampling procedures have been used. Nowadays, we have adopted the transabdominal approach because, in our experience, it seems to be associated with a low risk (2%) of fetal mortality. At the present time, the beta-thalassemia mutations are detected directly by dot-blot analysis of amplified DNA with 32P- or horseradish peroxidase-labeled allele-specific oligonucleotide probes. Two oligonucleotide probes, one complementary to the codon-39 nonsense mutation, which accounts for 95.7% of the beta-thalassemia chromosomes in the Sardinian population, and the other complementary to the frameshift at codon 6, which is the second most common mutation in our population (2.1%), allow us to make prenatal diagnosis in the large majority of cases. Notwithstanding a careful dissection of maternal decidua from chorionic villi, co-amplification of maternal sequence was detected in 4 out of 425 cases tested by this procedure. In order to avoid this pitfall, the simultaneous amplification of highly polymorphic VNTR (variable number of tandem repeats) segments could be used. On the whole we have so far carried out 2711 prenatal tests: 1130 by fetal blood analysis, 1156 by oligonucleotide hybridization on electrophoretically separated DNA fragments, and 425 by dot-blot analysis on amplified DNA with allele-specific oligonucleotide probes. Two errors occurred by fetal blood analysis and none by DNA analysis. The incidence of thalassemia major declined from 1:250 live births in the absence of prevention to 1:1000 after the establishment of this program, indicating that carrier screening and prenatal diagnosis are effective means for preventing thalassemia major at the population level.

Nuchal translucency and the acceptance of invasive prenatal chromosomal diagnosis in women aged 35 and older.

OBJECTIVE: To investigate the effect of nuchal translucency screening on use of prenatal diagnosis for chromosomal abnormalities in women aged 35 and older. METHODS: Two groups of women, referred to our center for prenatal karyotype diagnosis because of maternal age, were compared: one in 1995 and the other in 1999 after the introduction of nuchal translucency measurement. Each woman received nondirective genetic counseling, and for the 1999 group, nuchal translucency results were also discussed. Risks of transabdominal chorionic villi sampling (CVS) and amniocentesis, laboratory techniques, genetic results, and local experiences were discussed. Patient's decision to undergo prenatal diagnosis, acceptance of the nuchal translucency test (in the 1999 group), and the rate of chromosomal abnormalities diagnosed by transabdominal CVS and amniocentesis, were considered. RESULTS: Two hundred twenty-one of 982 (22%) women in the 1995 group and 421 of 1386 (30%) in the 1999 group, after nondirective genetic counseling declined invasive diagnosis (P <.05). In the 1999 cohort, 1088 of 1089 (99.9%) women of appropriate gestational age had nuchal translucency measurement. Among women seen in 1995, 214 opted for transabdominal CVS (31%) and 476 (69%) for amniocentesis. Nineteen abnormal karyotypes were detected, six by transabdominal CVS and 13 (68.5%) by amniocentesis. In 1999, 266 women (29%) opted for transabdominal CVS and 650 (71%) for amniocentesis. Twenty abnormal karyotypes were detected, 13 (65%) by transabdominal CVS and seven (35%) by amniocentesis (P <.05). CONCLUSION: Knowledge of nuchal translucency could lead to a decrease in the demand for invasive diagnosis and to a more frequent diagnosis by first-trimester transabdominal CVS.

A study on the distribution of Bothriocephalus andresi (Cestoda, Pseudophyllidea) in Citharus linguatula.

The distribution of Bothriocephalus andresi (Porta, 1911) in a population of Citharus linguatula (L.) caught off the Tuscan coasts was studied for one year. The negative binomial model proved useful for analysing the distribution of the parasite (B. andresi) in the host population. The ability of the parasite to adjust not only to its own environment but also to that of its host was clear, as it may be evinced from the greater amount of eggs and, hence, of larvae noticed in the period when there was abundance of plankton (high number of intermediate hosts). The balance achieved between host and parasite populations was also evident, and the modulation of the intensity of the infestation was such that it may be imputed to a greater immune response in the host.

[Fertilization in vitro and microinsemination. 6 years of experience in the Ospedale Microcitemico of Cagliari]. / Fecondazione in vitro e microinseminazione. Sei anni di esperienza nell'Ospedale Microcitemico di Cagliari.

BACKGROUND: In this study the results of six years experience (1993-1998) in IVF, ICSI and assisted hatching on 442 sterile couples for a total of 868 cycles are reported. Since 1997 ICSI has also been carried out in cases of azoospermia extracting mobile spermatozoa from the epididymal (MESA) or from the testicle (TESE). METHODS: Ages ranged from 20 to 48 and mean years sterility was 5.868 cycles were carried out of which 153 (17.62%) were for IVF, 705 (81.22%) for ICSI and 10 (1.15%) for MESA and TESE. Assisted hatching was performed through "partial zona dissection" in 329 cases on a total of 987 embryos. RESULTS: The pregnancy rates per embryo transfer in IVF and ICSI cycles were 22.4 and 19.96% respectively. Results considering patients age were: 33.87% below 35, 29.55% between 35 and 38 and 6.60% above 38. In the group of 189 assisted hatching patients the evolutive pregnancy rate rose from 15.78 to 26.40% and multiple pregnancies from 28.63 to 42%. CONCLUSIONS: ICSI has offered high rates of fertilization and pregnancy even in extreme cases of oligoasthenospermia or cases of azoospermia adopting MESA or TESE techniques. A decisive factor on pregnancy rates is age, very low over 38 years. Assisted Hatching further increased the pregnancy rates.

Intracardiac Doppler assessment of left valve inflow in first-trimester fetuses with increased nuchal translucency: preliminary observations in trisomy 21.

OBJECTIVES: To perform a qualitative assessment of the pulsed Doppler waveform profile at the level of left atrioventricular valve inflow in first-trimester fetuses with increased nuchal translucency thickness (NT), in order to compare those with trisomy 21 and those with normal karyotype. METHODS: This was a review of 285 consecutive fetuses with increased NT. Pulsed Doppler velocity waveforms of left atrioventricular valve inflow were recorded. The E-wave, A-wave and velocity profile in the aorta were displayed. Cases were classified into two patterns: Pattern A included those in which the E-wave velocity crossed the A-wave before the baseline in all waveforms; Pattern B included those in which the lowest E-wave velocity crossed the baseline but not the A-wave in at least one of the profiles. The karyotype was determined and the frequency of occurrence of Patterns A or B in fetuses with normal karyotype and those with trisomy 21 were compared. RESULTS: Of the 285 cases, 230 were assigned to Pattern A and 55 to Pattern B. There were 47 cases of trisomy 21, 22 had other chromosomal abnormalities, and 212 had a normal karyotype; in four cases the karyotype was unknown. Among the 212 karyotypically normal fetuses, five had heart defects, five had other structural defects, three suffered spontaneous intrauterine death and one was terminated. Pattern A was found in 200/212 (94.3%) cases with normal karyotype, in 12/47 (25.5%) cases with trisomy 21, and in 17/22 (77.3%) cases with other chromosomal abnormalities. Pattern B was found in 12/212 (5.7%) cases with normal karyotype, in 35/47 (74.5%) cases with trisomy 21 (chi-square test, P < 0.001), and in 5/22 (22.7%) cases with other chromosomal abnormalities. CONCLUSIONS: Intracardiac Doppler qualitative assessment of left valve inflow in first-trimester fetuses with increased NT shows differences between normal and trisomy 21 fetuses, probably reflecting differences in myocardial function.

Antigenicity of Pseudodactylogyrus anguillae and P. bini (Monogenea) in the European eel (Anguilla anguilla, L.) under different oxygenation conditions.

The antibody response of European eels (Anguilla anguilla, L.) to the branchial parasites Pseudodactylogyrus anguillae and P. bini under hyperoxygenation conditions was studied. The antigenic fractions of parasites were detected by means of electrophoretic techniques (SDS-PAGE) and by Western blot analysis. The results obtained demonstrate that under hyperoxygenation conditions, the eels responded to a greater number of proteins, and this was correlated with a decrease in the level of infestation.

Umbilical artery velocity waveforms before and after chorionic villus sampling.

Because a vascular aetiology has been suggested for the limb and oromandibular defects described after chorionic villus sampling (CVS), to determine whether transabdominal (TA) CVS causes noticeable changes in umbilical artery velocity waveforms in first-trimester pregnancies, the pulsatility index (PI) of the umbilical artery was evaluated before and after TA-CVS in 175 pregnancies sampled between 10.0 and 13.0 weeks' gestation. In 139 uncomplicated pregnancies, the mean PI values (with 95 per cent confidence interval) were before TA-CVS 2.751 (2.692-2.809), after 10 min 2.723 (2.697-2.809), and after 1 h 2.781 (2.722-2.840). There were no significant changes in PI relative to the CVS procedure either in pregnancies with an abnormal result or in those ending in spontaneous abortion. Our data do not support any statistically significant change in umbilical artery PI relative to TA-CVS in first-trimester pregnancies. This procedure, despite its invasive character, does not appear to affect the feto-placental circulation.

Chorionic villus sampling and acceptance rate of prenatal diagnosis.

In this paper, we compared the acceptance rate of fetal diagnosis for beta-thalassemia in three group of couples of Sardinian descent; the first counselled before DNA analysis was available, the second presenting after DNA analysis was introduced but too late for chorionic villus sampling and thus monitored by amniocyte DNA analysis and the third presenting within the first trimester after DNA analysis was introduced and thus in time for trophoblast DNA analysis. A higher proportion of couples from the latter group opted for fetal testing as compared to the 1st and 2nd group. These results indicate that in this population, introduction of 1st trimester diagnosis made prenatal testing acceptable to practically all counselled couples at risk.