White blood cell count: a valuable tool for suspecting Cushing's syndrome.

PURPOSE: Simple screening tests to determine whether Cushing's syndrome (CS) should be ruled out are lacking. Tools that enable early diagnosis could reduce morbidity and associated sequelae. The potential of glucocorticoid-induced changes in the white blood cell (WBC) count for raising suspicion of CS is assessed. METHODS: This was a retrospective case‒control study. The WBC counts of 73 cases with CS and 146 matched controls were compared. The number of leukocytes (Leu), the number and percentage of neutrophils (N, Np), the number and percentage of lymphocytes (L, Lp), neutrophil-to-lymphocyte differences in the number and percentage (N-L, Np-Lp), neutrophil-to-lymphocyte ratio in the number and percentage (NLR, NLRp), and leukocyte-to-lymphocyte differences (Leu-L) were evaluated. The area under the ROC curve (AUC) was calculated for each of these parameters. Reference values were estimated that could help disclose occult CS. RESULTS: All ten parameters showed significant differences between cases and controls. The AUC was greater than 0.7 for all ten parameters, and was the best for the NLRp and Lp (AUC: 0.89). An Lp of 23.9% showed a diagnostic accuracy of 84.9% for the diagnosis of CS. The concordance of an Lp below 24% and more than 8000 leucocytes had a PPV of 78.2% for CS, while the pairing of an Lp over 24% and a Leu below 8000 cells had an NPV of 97.3% for CS. CONCLUSION: WBC count assessment can be an effective tool to raise suspicion of CS, prompting diagnostic testing. This simple and universally available test may allow earlier diagnosis of CS before highly evolved phenotypes develop.

Selecting optimal R6TX2 intermetallics (R = Gd, Tb, Dy; T = Mn, Fe, Co, Ni; X = Sb, Te) for magnetic refrigeration.

A complete experimental study of the physical properties playing a relevant role in the magnetic refrigeration application (structural, magnetic, magnetocaloric and thermal) has been performed over nine selected Fe2P-type R6TX2 (R = Gd, Tb, Dy; T = Mn, Fe, Co, Ni; X = Sb, Te) intermetallic compounds, to work close to room temperature. Two magnetic phase transitions are observed for these materials: a paramagnetic to ferromagnetic transition in the range of 182-282 K and a spin reorientation transition in the range of 26-76 K. As a consequence, two peaks related to a direct magnetocaloric effect (DMCE) appear with the magnetic entropy change, generating a wide table-like plateau region in between both peaks, which is required to improve the efficiency of refrigerators following an Ericsson cycle. The highest magnetic entropy peak value for µ0ΔH = 5 T is found for Tb2Dy4FeSb2, with 7.72 J kg-1 K-1 around 182 K. For the same applied field the other compounds show moderate values around room temperature (2.88-4.53 J kg-1 K-1). However, the superposition of the two peaks results in huge refrigerant capacity values, up to RCFWHM(5 T) = 1103.04 J kg-1 in the case of Tb2Dy4FeSb2. The thermal diffusivity, thermal effusivity, thermal conductivity and specific heat capacity have been measured at room temperature, and the temperature dependence of the former has been obtained around the relevant magnetic phase transition region, with values in the range of 1.3-2.3 mm2 s-1, which are good for magnetic refrigerators at high working frequencies. The study is completed with a rigorous critical behavior analysis of the second order PM-FM transition. The critical exponent γ points to long range order interactions, in general, while ß values are in the range of 0.59-0.90, indicating a deviation from theoretical models as a reflection of the magnetic complexity in these compounds. The critical exponents have been used to confirm the scaling relations of magnetocaloric properties, and the scaling of refrigerant capacity (RC) values in materials exhibiting two magnetic phase transitions is addressed, concluding that for a correct scaling of RC the magnetic entropy change peak must be considered symmetric. The role of each atom in the properties of the compounds is discussed.

Critical behavior near the Lifshitz point in Sn(2)P(2)(S(1 - x)Se(x))(6) ferroelectric semiconductors from thermal diffusivity measurements.

The thermal diffusivity of the ferroelectric family Sn(2)P(2)(Se(x)S(1 - x))(6) (0 ≤ x ≤ 1) has been measured by a high-resolution ac photopyroelectric technique, using single crystals, with the aim of studying the evolution of the ferroelectric transition with Se doping. Its change from second order character to first order while passing the Lifshitz point (x approximately 0.28) has been evaluated, as well as the splitting of the transition at high Se concentrations. The critical behavior of the ferroelectric transition in terms of the different universality classes and their underlying physical dominant effects (tricriticality, long-range dipole interactions, Lifshitz point) has been discussed using thermal diffusivity measurements in the very close vicinity of the critical temperature. This study reveals that for Se concentrations around the Lifshitz point, long-range dipole interactions do not play a significant role and that the critical parameters are close to those predicted for the Lifshitz universality class.

Lack of evidence of paratuberculosis in wild canids from Southwestern Europe.

Wild carnivores are at the top of the trophic chain. They are predators and carrion consumers, and thus, prone to come in contact with disease agents contaminating the environment or infecting live or dead animals. We hypothesized that wild canids could be used as sentinels for the detection of regions with higher Mycobacterium avium paratuberculosis (MAP) prevalence in wild and domestic animals. To test this hypothesis, we set up an ELISA to test 262 wolf (Canis lupus) and fox (Vulpes vulpes) sera for MAP-specific antibodies and processed a subset of samples for culture (n = 61), MAP-specific PCR (15) and histopathology (14). In wolves, the optical density (OD) values in the ELISA were continuously distributed. Ten fox sera (4%) had OD readings of over twice the mean, suggesting contact with mycobacteria. However, all samples tested by PCR were negative for both IS900 and ISMAP02 sequences, and samples cultured for MAP yielded no growth. No visible paratuberculosis or tuberculosis-compatible lesions were recorded. On histopathological examination, no lesions compatible with mycobacterial diseases were observed. These results suggest that wild canids show little or no evidence of paratuberculosis and are unlikely to be useful sentinels for the detection of MAP in Southwestern Europe.

Results of a nationwide survey on multidisciplinary teams on thyroid cancer in Spain.

AIM: There is an important lack of knowledge as to the functioning of multidisciplinary teams on thyroid cancer in current clinical practice. We aimed to retrieve data on the composition, structure, and procedures developed by the multidisciplinary units of thyroid cancer in Spain. METHODS: A nationwide survey consisting of questions about composition, structure, and functioning of multidisciplinary teams was designed. It was available online from November 15, 2017 to February 15, 2018. RESULTS: Seventy-two multidisciplinary units responded to our survey. Of these, 15 (20.8%) focused only in thyroid cancer, while 57 (79.2%) included other endocrine disorders or non-endocrine tumors. The median (interquartile range) of members of the teams was 11 (9-14). The most frequent medical specialties in the units were endocrinology (100%), surgery (94.4%), pathology (80.6%), radiology (75.0%), nuclear medicine (73.6%), and medical oncology (55.6%). The annual number of patients reviewed by the teams was 40 (20-74). 56.9% of the multidisciplinary teams have elaborated clinical protocols for local use. Apart from clinical case discussions in the meetings, 45.8% of the units included educational activities and 36.1% research subjects. Quality indicators were developed by 22% of the teams. CONCLUSIONS: These results suggest that there are some hopeful signs that international recommendations of having multidisciplinary approach to patients with thyroid cancer are being followed in Spain. This gives us the opportunity to proceed with further studies to analyze the real impact of this high standard of care on patient outcomes.

Sarcoptic mange in red deer from Spain: improved surveillance or disease emergence?

Concern about emerging diseases has risen in recent years, and multihost situations have become increasingly relevant for wildlife management and conservation. We present data on Asturias, northern Spain, where 80 mangy red deer (Cervus elaphus) have been found since the beginning of the epizootic in chamois (Rupicapra pyrenaica parva) in 1993. We combine field and necropsy data with the results of a serosurvey using an in-house ELISA test to evaluate if deer mange due to Sarcoptes scabiei is an emerging disease in this area. The mean number of deer mange cases per year was 5, with a maximum of 16. No significant relationship was detected between monthly temperatures, rainfall or number of days with snow cover and the annual number of sarcoptic mange cases in red deer. Only 4 mangy red deer (5%) were detected outside the limits of scabietic chamois distribution during the same year, and all were less than 2500 m away from that limit. The longest distance reported between two consecutive mangy deer locations was 18 km. Mange cases were significantly more frequent in stags than in hinds and in adults than in juvenile deer. The time of the first mange detection in chamois in each sector, year with minimum number of chamois recorded, year with maximum chamois population decline rate and chamois density offered no significant correlation with red deer mange cases appearance moment and frequency. In the mange affected area, ELISA testing of 327 blood samples from hunter-harvested deer without obvious mange-compatible lesions revealed only 4 seropositive animals. All 83 sera from hunting preserves without clinical cases yielded negative ELISA results. According to these epidemiological data mange does not seem to threaten red deer populations in Asturias. However, continued monitoring of deer health and ELISA testing for sarcoptic mange is advisable.

Characterization of widespread canine leishmaniasis among wild carnivores from Spain.

Visceral Leishmaniasis (VL) is an emerging zoonotic parasitic disease caused by Leishmania infantum in Mediterranean countries, with sand flies (Phlebotomus spp.) as vectors and dogs as the main domestic reservoir. The role of wild carnivores in the epidemiology of leishmaniasis is still controversial. In order to determine the prevalence of natural infection with L. infantum in wild carnivores from Spain, we analyzed 217 samples by PCR and western blotting and used restriction fragment length polymorphism (RFLP) to compare the patterns present in wild carnivores with those of domestic dogs from the same areas. DNA of the parasite was detected in spleen or blood samples from 35 (16.12%) analyzed wild carnivores, including 8 of 39 (20.5%) wolves (Canis lupus), 23 of 162 (14.1%) foxes (Vulpes vulpes), 2 of 7 (28.6%) Egyptian mongooses (Herpestes ichneumon), 1 of 4 genets (Geneta geneta), and 1 of 4 Iberian lynxes (Lynx pardinus). No significant sex or age differences in prevalence were observed in wolves and foxes (P>0.05), but there was a significant difference among regions in foxes (P<0.05). A total of 12 PCR-RFLP patterns were found in foxes, 6 in wolves, 4 in dogs, 2 in Egyptian mongooses and 1 in lynx and genet. RFLP patterns differed between dogs and foxes in the two areas where they could be compared. This is the first study of canine leishmaniasis in wild canids and other carnivores from different regions of Spain by PCR. The prevalence of infection indicates the existence of natural infection in apparently healthy wild carnivore populations, and our results are suggestive of a sylvatic cycle independent of dogs.

Acaricidal activity of fluralaner against Ornithodoros moubata and Ornithodoros erraticus argasid ticks evaluated through in vitro feeding.

Ornithodoros erraticus and Ornithodoros moubata are argasid tick vectors that transmit severe diseases such as African swine fever and human relapsing fever. Elimination of the synanthropic populations of these vectors would facilitate the control of these diseases. Fluralaner is a novel isoxazoline that selectively blocks the GABA- and glutamate-gated channels, providing potent insecticidal and acaricidal activity. The aim of the current study was to provide quantitative data on the susceptibility of males, females and third nymphal instar of O. erraticus and O. moubata to fluralaner through in vitro feeding exposure. Fluralaner activity against these developmental stages and species was assessed by feeding the ticks on ovine blood medicated with decreasing fluralaner concentrations between 1 and 10-8µg/mL. Tick mortality was measured at 4, 24 and 48h and 1, 2 and 3 weeks post-feeding. Tests included solvent-treated and untreated blood controls. Fluralaner was extremely active against O. erraticus, with mean lethal concentrations 50 (LC50) and 95 (LC95) of 2.0×10-8 and 5.4×10-8µg/mL, respectively. Fluralaner was also highly active against O. moubata, showing a mean LC50 of 1.5×10-6µg/mL and a mean LC95 of 1.8×10-3µg/mL. In the latter species, the most susceptible life stages were the females (LC95 1.4×10-4µg/mL). Fluralaner demonstrated potent acaricidal activity against all developmental stages of O. erraticus and O. moubata tested, in the first 48h after in vitro feeding. Therefore, fluralaner has the potential to provide very high acaricidal efficacy to multiple argasid tick species via feeding exposure and could be included as an acaricidal agent in integrated programmes for the control of argasid tick vectors and argasid tick-borne diseases.

Serological Surveillance and Direct Field Searching Reaffirm the Absence of Ornithodoros Erraticus Ticks Role in African Swine Fever Cycle in Sardinia.

African swine fever (ASF), one of the most important diseases of swine, has been endemic in the Italian island of Sardinia for more than 35 years. During these decades, several strategies and eradication efforts have been implemented in the island with limited success. Strong climatic and ecological similarities exist between Sardinia and one area of the Iberian Peninsula where Ornithodoros erraticus ticks were involved in the persistence of ASF from 1960 to 1995. This fact leads to the hypothesis that, potentially, Ornithodoros ticks could be also involved in the ASF cycle in Sardinia, thus accounting for some of the reoccurring ASF outbreaks in this island. Initial efforts aimed at detection of Ornithodoros ticks in Sardinia were performed during the 1970s/1980s with no positive results. Accordingly, the absence of Ornithodoros ticks in Sardinia has been generally accepted. However, since a new and reinforced ASF eradication programme has been recently launched in Sardinia, it is essential to clarify the presence and role of these soft ticks in the epizootiology of ASF in this island. For that purpose, 1767 porcine serum samples collected from all around the island (1261 from domestic and 506 from wild boar) were analysed by ELISA for antibodies to salivary antigens of Ornithodoros erraticus. In addition, Ornithodoros ticks were directly searched in a number of pig premises that have suitable habitats for these ticks and were located in areas repeatedly affected by ASF. Only one serum sample resulted positive in the serological assay, and no Ornithodoros ticks were collected in none of the premises. These results indicate that these soft tick species are not involved in the epizootic cycle of ASF in Sardinia and highlight the importance of controlling other risk factors still present in the island for effectively eradicate the disease.

TSGP4 from Ornithodoros moubata: molecular cloning, phylogenetic analysis and vaccine efficacy of a new member of the lipocalin clade of cysteinyl leukotriene scavengers.

Recently obtained evidence indicated that an orthologue of the O. savignyi TSGP4 salivary lipocalin was present in the saliva of O. moubata. TSGP4 is known to act as a cysteinyl leukotrienes scavenger helping in the prevention of inflammation and oedema at the tick bite site. Since this function seems to be crucial for successful tick feeding, the novel O. moubata TSGP4 turned into a potential vaccine target. The purposes of the current work were: (i) to clone and characterize the O. moubata TSGP4 and, (ii) to produce it as recombinant to evaluate its protective efficacy as vaccine antigen. The results of these experiments indicated that the O. moubata TSGP4 shows high sequence and structural identity with the O. savignyi orthologue suggesting identical function in the physiology of the tick-host relationship. The mature native TSGP4 is not immunogenic when it is inoculated to host with tick saliva during feeding, but host vaccination with the recombinant protein TSGP4 in Freund's adjuvants induced strong humoral immune responses that recognized both the recombinant and native TSGP4 and protected the host with a 14.1% efficacy. So, the O. moubata TSGP4 can be considered a silent salivary antigen; however, in the light of the current results, its inclusion in the current repertory of protective antigens to be targeted by anti-tick vaccines could be controversial.

Short-term safety, tolerability and efficacy of a very low-calorie-ketogenic diet interventional weight loss program versus hypocaloric diet in patients with type 2 diabetes mellitus.

UNLABELLED: Brackground:The safety and tolerability of very low-calorie-ketogenic (VLCK) diets are a current concern in the treatment of obese type 2 diabetes mellitus (T2DM) patients. OBJECTIVE: Evaluating the short-term safety and tolerability of a VLCK diet (<50 g of carbohydrate daily) in an interventional weight loss program including lifestyle and behavioral modification support (Diaprokal Method) in subjects with T2DM. METHODS: Eighty-nine men and women, aged between 30 and 65 years, with T2DM and body mass index between 30 and 35 kg m(-)(2) participated in this prospective, open-label, multi-centric randomized clinical trial with a duration of 4 months. Forty-five subjects were randomly assigned to the interventional weight loss (VLCK diet), and 44 to the standard low-calorie diet. RESULTS: No significant differences in the laboratory safety parameters were found between the two study groups. Changes in the urine albumin-to-creatinine ratio in VLCK diet were not significant and were comparable to control group. Creatinine and blood urea nitrogen did not change significantly relative to baseline nor between groups. Weight loss and reduction in waist circumference in the VLCK diet group were significantly larger than in control subjects (both P<0.001). The decline in HbA1c and glycemic control was larger in the VLCK diet group (P<0.05). No serious adverse events were reported and mild AE in the VLCK diet group declined at last follow-up. CONCLUSIONS: The interventional weight loss program based on a VLCK diet is most effective in reducing body weight and improvement of glycemic control than a standard hypocaloric diet with safety and good tolerance for T2DM patients.

Concomitance and interactions of pathogens in the Iberian wolf (Canis lupus).

With the aim of improving our understanding of their epidemiological features, exposure to or presence of Canine Parvovirus (CPV), Canine Distemper Virus (CDV), Leishmania infantum and Sarcoptes scabiei were studied in 88 wild wolves from Asturias (Northern Spain) by means of long-term (2004-2010) serological and molecular data. Individual and population factors and the possible interactions between them were also statistically analyzed for better understanding the contact/presence of studied pathogens. The overall seroprevalence values were 19%, 61%, 20% and 0% for CDV, CPV, S. scabiei and Leishmania, respectively, while a 46% of studied wolves showed Leishmania genetic material presence. Sarcoptic mange, CDV and CPV showed higher seroprevalence values in the areas with higher wolf densities, and a positive association between CDV and S. scabiei antibody responses was detected. Reported data highlight the need of considering concomitant pathogens and their possible interactions for a better understanding of diseases and their management in wildlife.

Vaccination of sheep against Fasciola hepatica with homologous fatty acid binding proteins.

The current study was designed to test the immunoprophylactic properties of native (nFh12) and recombinant (rFh15) antigens from Fasciola hepatica in sheep subsequently infected with the fluke. Thirty lambs were divided into six groups according to various patterns of immunisation and times of infection and necropsy. The antigens were emulsified in Freund's adjuvant. Levels of specific anti-nFh12 and anti-rFh15 antibodies rose rapidly by 2 weeks after the first immunisation and were always significantly higher in immunised-infected sheep than in control-infected sheep. On completion of the trial there was no difference in fluke burden between groups vaccinated with either of the antigens and non-immunised controls. However, worm size and faecal egg counts were significantly diminished in the sheep vaccinated with either of the antigens, suggesting an anti-fecundity effect. This is the first report of experimental vaccination of sheep against F. hepatica with purified native and recombinant antigens related to fatty acid binding proteins.

Purification, N-terminal sequencing and diagnostic value of the major antigens of Ornithodoros erraticus and O. moubata.

To enhance the specificity and sensitivity of serological detection of swine exposed to Ornithodoros erraticus or O. moubata, we purified the 158, 186, 215 and 260 kDa antigens from the former species and the designated (owing to their MW and charge) 19C, 17A, 20A1 and 20A2 antigens of the latter by HPLC and gel electroelution methods. All the O. erraticus antigens share epitopes and are difficult to purify individually by reverse phase and ion-exchange chromatography due to their molecular similarity. Tested individually by ELISA, all of them give the same optical densities (OD) with anti-O. erraticus sera, and these ODs are always lower with anti-immature than with anti-adult sera. Although immature and adult specimens have the same antigens, immature forms induce more anti-carbohydrate antibodies than adults. This is the reason for the lower ODs of the anti-immature sera against purified antigens, since these latter antigens essentially react with anti-peptide antibodies (hence, increasing the specificity and sensitivity of the serology). The N-terminus of the 260 kDa antigen shows 80-90% similarity with the hemoglobin alpha-chain of many mammals. The antigens of O. moubata are proteins that are very different from one another and are, therefore, readily purified by ion exchange chromatography. The 20A1 antigen appears to be the most immunogenic and is recognized equally by anti-immature and anti-adult sera. This antigen does not give false positive reactions with the negative control sera analyzed and its N-terminus region shares 46.2% homology with the alpha-chain of the C3 component of rabbit complement.

A fatty acid binding protein from Fasciola hepatica induced protection in C57/BL mice from challenge infection with Schistosoma bovis.

Three strains of mice (NMRI, C57/BL, BALB/c) were each immunized with a 12 kDa purified, native Fasciola hepatica fatty acid binding protein (Fh12) and challenged percutaneously with Schistosoma bovis cercariae. C57/BL mice immunized with Fh12 had significant reductions in S. bovis worm burden recoveries (96 and 87% reductions over controls in two separate experiments). When using NMRI or BALB/c mice, Fh12 alone or in Freund's adjuvant failed to induce significant protection against S. bovis. In C57/BL mice vaccinated against Fh 12, antibodies to the IgG2a isotype, but not to the IgG1 isotype, increased by 2 weeks after the second immunization and remained high through 8 weeks of S. bovis infection. Antibodies to S. bovis increased after 4 weeks of infection. Regarding cytokine production by spleen mononuclear cells, C57/BL mice vaccinated with Fh12 in adjuvant, and having the highest protective response against challenge infection with S. bovis, had an increase of IFN-gamma production with Concanavalin A but no increase of IL-4 in similarly stimulated cells. These results suggest that the protection obtained in this group of mice is mediated by a Th1 immune response.

Evaluation of an enzyme-linked immunosorbent assay to detect specific antibodies in pigs infested with the tick Ornithodoros erraticus (Argasidae).

Ornithodoros erraticus is known to transmit the virus that causes the highly contagious disease, African Swine Fever, in Spain. As part of the disease eradication campaign, an ELISA test to detect specific antibodies against the tick was developed. The ELISA, using salivary gland preparations as an antigen, showed high sensitivity and was able to detect as few as 10 adult ticks. The specific antibodies were detected in the sera 6 weeks after the primary infestation and strongly increased after the challenge. The utility of this test under field conditions was also tested.

Vaccination of mice against schistosoma bovis with a recombinant fatty acid binding protein from Fasciola hepatica.

Two strains of mice (NMRI and C57/BL) were each immunized with a 15kDa recombinant Fasciola hepatica fatty acid binding protein (FABP) (Fh15) and challenged percutaneously with Schistosoma bovis cercariae. C57/BL mice immunized with Fh15 had significant reductions in S. bovis worm burden recoveries (72% reductions over controls). When using NMRI mice, Fh15 in Freund's adjuvant failed to induce significant protection against S. bovis. In C57/BL mice, only antibodies to the IgG2a isotype increased after the second immunization and remained high through 8 weeks of S. bovis infection. This is the first time that a heterologous recombinant molecule from F. hepatica has been used in vaccination against S. bovis, obtaining a significant reduction in the number of worms in C57/BL mice.

Vaccination of mice and sheep with Fh12 FABP from Fasciola hepatica using the new adjuvant/immunomodulator system ADAD.

We evaluate the ability of a Fasciola hepatica FABP native antigen (Fh12) with a new vaccination system called ADAD to protect mice and sheep against an experimental F. hepatica infection. The vaccination protocol consists of a set of two injections. The first injection contains a micelle in which two components are included, saponin from Quillaja saponaria (Qs) and/or Anapsos (A) a Polypodium leucotomos hydroalcoholic extract, both emulsified in a non-mineral oil (Montanide) in a water/oil emulsion (30/70). This is subcutaneously injected to achieve the "adaptation" of the immune system to subsequent stimuli. The second injection contains in addition the Fh12 antigen. Two different experiments were carried out using two mouse strains (BALB/c and CD-1). Mice vaccinated with Qs+A+Fh12 presented a survival rate of 40%, when compared with control groups. Furthermore, we evaluated the efficiency of the vaccination in sheep against an experimental F. hepatica challenge. The vaccinated sheep presented lower fluke recovery (24.5%), number of eggs in bile fluid (58.1%) and faeces (40.3%) than control groups. The recovered flukes were shorter (32.7%), immature (34.0%) and with lower body mass (31.6%) than non-complete vaccinated sheep. Thus, the new ADAD system could be a good alternative for future vaccination experiments against fasciolosis.

Relationships between the defensive systems of Iberian-breed swine and the European vector of African swine fever, Ornithodoros erraticus.

To discover whether the immune system of Iberian-breed pigs exerts any adverse action on Ornithodoros erraticus, 3 pairs of pigs were subjected to a weekly infestation over 12 wk with 1,000 larvae, 500 nymphs-1, or 200 adults. Each pair was bitten by only 1 developmental stage. Batches of parasites identical to the foregoing ones were fed weekly on control swine. In none of the 10 parameters studied for each of the batches fed weekly was any significant difference found that could be attributed to the state of sensitization of the animals in which, in a previous study, the presence of high titers of anti-O. erraticus antibodies was found. It was observed that the possible pruritus due to immediate hypersensitivity reactions, which in the test animals appeared after the third week, had no protective value in the natural milieu. In view of the inability of the swine to exert any control over the soft ticks, it is concluded that the size of their populations in the pig pens and their composition according to the developmental stage are factors that depend exclusively on the opportunities that swine breeders offer such populations to feed on the animals.

Immunoprophylaxis against Fasciola hepatica in rabbits using a recombinant Fh15 fatty acid-binding protein.

Previous studies of ours have demonstrated that a recombinant protein (Fh15) related to fatty acid-binding proteins did not induce significant protection in rabbits challenged 2 or 4 wk postimmunization over nonimmunized controls. In the current study, rabbits were immunized with Fh15 and challenged with Fasciola hepatica metacercariae 12 and 20 wk later. In the current study in which longer lag periods for challenge infection after the second immunization were used, worm burden reductions compared to adjuvant controls were a significant 43% and 76%, respectively. Importantly, rabbits immunized with Fh15 had significant numbers of immature flukes, 66% in the 12-wk period and 84% in the 20-wk lag period as compared to controls. In addition, liver lesions were clearly diminished in the vaccinated rabbits. Enzyme-linked immunosorbent assay absorbance values showed that immunized rabbits developed high antibody levels to Fh15 from 8 wk after the first immunization and did not increase after challenge. These results suggest that a recombinant F. hepatica molecule related to fatty acid-binding proteins induces protective (worm burden reductions), anti-fecundity (immature flukes), and anti-pathology (less liver lesions) effects in rabbits and may serve as a model for the immunoprophylaxis of fascioliasis.