Scalp eschar and neck lymphadenopathy by Rickettsia slovaca after Dermacentor marginatus tick bite case report: multidisciplinary approach to a tick-borne disease.

BACKGROUND: Scalp Eschar and Neck LymphAdenopathy after Tick bite is a zoonotic non-pathogen-specific disease most commonly due to Rickettsia slovaca and Rickettsia raoultii. Diagnosis is mostly based only on epidemiological and clinical findings, without serological or molecular corroboration. We presented a clinical case in which diagnosis was supported by entomological identification and by R. slovaca DNA amplifications from the tick vector. CASE PRESENTATION: A 6-year-old child presented with asthenia, scalp eschar and supraclavicular and lateral-cervical lymphadenopathy. Scalp Eschar and Neck LymphAdenopathy After Tick bite syndrome following a Dermacentor marginatus bite was diagnosed. Serological test on serum revealed an IgG titer of 1:1024 against spotted fever group rickettsiae, polymerase chain reaction assays on tick identified Rickettsia slovaca. Patient was successfully treated with doxycycline for 10 days. CONCLUSIONS: A multidisciplinary approach including epidemiological information, clinical evaluations, entomological identification and molecular investigations on tick, enabled proper diagnosis and therapy.

Donkey's Milk in the Management of Children with Cow's Milk protein allergy: nutritional and hygienic aspects.

BACKGROUND: The therapeutic strategy for children with cow's milk allergy (CMA) consists in the elimination of cow's milk (CM) from their diet. Donkey's milk (DM) has been reported to be an adequate alternative, mainly to his nutritional similarities with human milk (HM) and excellent palatability. The aim of present prospective study was to evaluate the nutritional impact of DM on the diet of children with CMA in term of children growth. METHODS: Before the nutritional trial on children and during the study the health and hygiene risks and nutritional and nutraceuticals parameters of DM were monitored. Children with CMA were identified by the execution of in vivo and in vitro tests for CM and subsequent assessment of tolerability of DM with oral food challenge (OFC). Finally, we prescribed DM to a selected group of patients for a period of 6 months during which we monitored the growth of children. A total of 81 children, 70 with IgE mediated cow's milk protein allergy (IgE-CMPA) and 11 with Food Protein Induced Enterocolitis Syndrome to CM (CM-FPIES), were enrolled. RESULTS: Seventy-eight out of 81 patients underwent the OFC with DM and only one patient with IgE-CMPA (1.5 %) reacted. Twenty-two out of 81 patients took part of the nutritional trial. All the 22 patients took and tolerated the DM, moreover DM did not change the normal growth rate of infants. CONCLUSIONS: In conclusion, DM resulted safe in term of health and hygiene risks and nutritionally adequate: no negative impact on the normal growth rate of children was assessed. Therefore, it may be a suitable alternative for the management of IgE mediated CMA and FPIES, also in the first 6 months of life, if adequately supplemented.

Amiata Donkey Milk Chain: Animal Health Evaluation and Milk Quality.

This study presents an investigation of Amiata donkey health and quality of milk for human consumption. Thirty-one lactating dairy jennies were examined. The following samples were collected: faecal samples from the rectum of animals for parasitological examination; cervical swabs for the detection of bacteria causing reproductive disorders; and blood samples for serological diagnosis of main zoonotic (Brucella spp., Leptospira spp.) and donkey abortion agents (Brucella spp., Leptospira spp., Salmonella abortus equi, Equine viral arterithis virus, Equine herpesvirus type 1). In addition, individual milk samples were collected and analysed for mastitis-causing pathogens and milk quality. Regarding animal health, we detected a high prevalence of strongyle parasites in donkeys. It is very important to tackle parasitic diseases correctly. Selective control programmes are preferable in order to reduce anthelmintic drug use. For dairy donkeys, withdrawal periods from anthelmintic drugs need to be carefully managed, in accordance with EU and national regulations. The isolation of Staphylococcus aureus in milk highlights the importance of preventing contamination during milking, by adopting appropriate hygiene and safety practices at a farm level. Lysozyme activity was high compared to cow's milk, contributing to the inhibitory activity against certain bacteria. Donkey milk was characterised by high lactose content, low caseins, low fat, higher levels of unsaturated fatty acids compared to ruminant milks. Unsaturated fatty acids and omega 3 fatty acids in particular have become known for their beneficial health effect, which is favourable for human diet. These characteristics make it suitable for infants and children affected by food intolerance/allergies to bovine milk proteins and multiple food allergies as well as for adults with dyslipidemias. It is also recommended to prevent cardiovascular diseases.

Indirect versus direct detection methods of Trichinella spp. infection in wild boar (Sus scrofa).

BACKGROUND: Trichinella spp. infections in wild boar (Sus scrofa), one of the main sources of human trichinellosis, continue to represent a public health problem. The detection of Trichinella spp. larvae in muscles of wild boar by digestion can prevent the occurrence of clinical trichinellosis in humans. However, the analytical sensitivity of digestion in the detection process is dependent on the quantity of tested muscle. Consequently, large quantities of muscle have to be digested to warrant surveillance programs, or more sensitive tests need to be employed. The use of indirect detection methods, such as the ELISA to detect Trichinella spp. infections in wild boar has limitations due to its low specificity. The aim of the study was to implement serological detection of anti-Trichinella spp. antibodies in meat juices from hunted wild boar for the surveillance of Trichinella spp. infections. METHODS: Two tests were used, ELISA for the initial screening test, and a specific and sensitive Western blot (Wb) as a confirmatory test. The circulation of anti-Trichinella IgG was determined in hunted wild boar muscle juice samples in 9 provinces of 5 Italian regions. RESULTS: From 1,462 muscle fluid samples, 315 (21.5%, 95% C.I. 19.51-23.73) were tested positive by ELISA. The 315 ELISA-positive muscle fluid samples were further tested by Wb and 32 (10.1%, 95% C.I. 7.29-13.99) of these were positive with a final seroprevalence of 2.2% (95% C.I 1.55-3.07; 32/1,462). Trichinella britovi larvae were detected by artificial digestion in muscle tissues of one (0.07%, 95%C.I. 0.01-0.39) out of the 1,462 hunted wild boars. No Trichinella spp. larvae were detected in Wb-negative wild boar. From 2006 to 2012, a prevalence of 0.017% was detected by muscle digestion in wild boar hunted in the whole Italian territory. CONCLUSIONS: The combined use of both serological methods had a sensitivity 31.4 times higher than that of the digestion (32/1,462 versus 1/1,462), suggesting their potential use for the surveillance of the Trichinella spp. infection in wild boar populations.

Development and Field Testing of a Real-Time PCR Assay for Caprine Arthritis-Encephalitis-Virus (CAEV).

Caprine arthritis/encephalitis (CAE) of goats and occasionally sheep are persistent virus infections caused by a lentivirus (CAEV). This viral infection results in arthritis in adult animals and encephalitis in kids. Prognosis for the encephalitic form is normally poor, with substantial economic loss for the farm. In this context an early/fast laboratory diagnosis for CAEV infection could be useful for effective prophylactic action. In this work we performed a quantitative real time PCR designed on the CAEV env gene to detect/quantify in goat/sheep samples, viral RNA or proviral DNA forms of CAEV. This procedure was validated in 15 sheep, experimentally infected with CAEV or with a highly correlated lentivirus (visna maedi, MVV); in addition, a total of 37 clinical goat specimens recruited in CAEV positive herds were analyzed and compared using serological analysis (Elisa and AGID). All samples infected with MVV resulted negative. In sheep experimentally infected with CAEV, proviral DNA was detectable 15 days post infection, whereas the serological methods revealed an indicative positivity after 40-60 days.This method showed a sensitivity of 10(2) env fragments/PCR) with a linear dynamic range of quantitation from 10(3) to 10(7)env fragments/PCR; the R2 correlation coefficient was 0.98. All subjects with a clinical diagnosis for Caprine Arthritis-Encephalitis (CAE) resulted CAEV DNA positive.

Flow cytometry analysis of immune cell populations isolated from cervicovaginal secretions of cynomolgus monkeys.

Flow cytometric analysis was used in this study to characterize the lymphocyte population present in the vaginal mucosa of the cynomolgus monkey. Vaginal immune cells were obtained, using absorbent wicks, from 11 normal cycling female monkeys at different stages of the menstrual cycle and from three nursing monkeys (not cycling). Leucocytes, including lymphocytes and monocyte-macrophage cells, were present in the cervicovaginal secretions of healthy cynomolgous primates throughout the three phases of the menstrual cycle. We also found that even if immune cells were constant throughout the menstrual cycle, among the T cell subsets there were differences. CD8+ cells [14.5+/-9% (mean+/-S.D.); range 3-30%] were more numerous compared to the mean number of CD4+ cells [7.3+/-5% (mean+/-S.D.); range 2-15%]. Characterization of the vaginal cells during the nursing period showed that the monocyte-macrophage (CD14+, CD11c+) cells were abundant compared with the low number of both B (CD20+) and T cells (CD2+). Our results show that cytometric analysis by FACS can be used to identify the immune cell populations present at the local level. This technique may provide a useful tool by which the vaginal environment can be studied in order to correlate cell phenotype with immune function.