Occurrence of atypical myxomatosis in Central Europe: clinical and virological examinations.

An outbreak of the atypical form of myxomatosis struck a rabbit farm in Hungary. The animals had previously been vaccinated with a vaccine containing Shope rabbit fibroma virus strain. The disease appeared in winter when the presence of mosquitoes and fleas is not common. The virus was isolated from an eyelid specimen of a naturally infected rabbit. The surviving animals were observed for four weeks, blood samples were collected and, after euthanasia, organ specimens were also examined by morphological methods including pathology and electron microscopy. Serum samples were examined by virus neutralisation for antibodies. Genetic analysis of the isolated virus was carried out by polymerase chain reaction (PCR) and direct sequencing. The primers were designed on the basis of the major envelope gene (Env) of the Lausanne reference strain in the GenBank. The viral proteins were examined by SDS-PAGE. The isolated virus (ref. no.: BP04/2001) was able to infect the susceptible animals directly, by contact. The disease was characterised by respiratory symptoms of the upper tracheal tract, conjunctivitis and high mortality by the 11th-14th day. Aerogenic infection with strain BP04/2001 resulted in 100% morbidity among the susceptible animals. Sequencing of the amplified 400-bp-long DNA revealed 97% homology with the Env gene of the Lausanne strain, which proves that strain BP04/2001 is a variant of the Lausanne strain having been enzootic throughout Europe. The live vaccine strain used in Hungary against myxomatosis, which is also a Lausanne-derived strain, protected the animals. According to the protein analysis a protein of 200 kDa in size is not expressed in strain BP04/2001. This is the first report on atypical myxomatosis in Central Europe. The virus spreads by airborne transmission and may cause severe losses in the rabbit population.

In vitro sensitivity of Hungarian Actinobaculum suis strains to selected antimicrobials.

In vitro antimicrobial sensitivity of 12 Hungarian isolates and the type strain ATCC 33144 of Actinobaculum suis to different antimicrobial compounds was determined both by the agar dilution and by the disc diffusion method. By agar dilution, MIC50 values in the range of 0.05-3.125 micrograms/ml were determined for penicillin, ampicillin, ceftiofur, doxycycline, tylosin, pleuromutilins, chloramphenicol, florfenicol, enrofloxacin and lincomycin. The MIC50 value of oxytetracycline and spectinomycin was 6.25 and 12.5 micrograms/ml, respectively. For ofloxacin, flumequine, neomycin, streptomycin, gentamicin, nalidixic acid, nitrofurantoin and sulphamethoxazole + trimethoprim MIC50 values were in the range of 25-100 micrograms/ml. With the disc diffusion method, all strains were sensitive to penicillin, cephalosporins examined, chloramphenicol and florfenicol, tetracyclines examined, pleuromutilins, lincomycin and tylosin. Variable sensitivity was observed for fluoroquinolones (flumequine, enrofloxacin, ofloxacin), most of the strains were susceptible to marbofloxacin. Almost all strains were resistant to aminoglycosides but most of them were sensitive to spectinomycin. A strong correlation was determined for disc diffusion and MIC results (Spearman's rho 0.789, p < 0001). MIC values of the type strain and MIC50 values of other tested strains did not differ significantly. Few strains showed a partially distinct resistance pattern for erythromycin, lincomycin and ampicillin in both methods.

Association between endometritis and urocystitis in culled sows.

Slaughterhouse sampling and examination of urogenital tracts of 499 sows and gilts culled for reproductive reasons from 21 Hungarian herds were performed over a 6-year period. The aim was to estimate the prevalence of different urogenital tract lesions, and to provide sensitivity and specificity estimates for macroscopic and bacteriological examinations in the diagnosis of urocystitis and endometritis. Furthermore, the association between endometritis and urocystitis was assessed. The prevalence of main lesions of the urogenital tract was similar to that reported in other studies. The 'sensitivity' of macroscopic and bacteriological methods was determined statistically by taking histopathology as the 'Gold Standard'. As a result, the 'sensitivity' of macroscopic methods for the diagnosis of endometritis and urocystitis was < or = 18.1% and 47.9%, respectively, while the 'sensitivity' of bacteriology for the diagnosis of the same conditions was < or = 31.8% and 63.0%, respectively. The presumed positive association between urocystitis and endometritis was confirmed; it was not confounded by parity. Animals affected by urocystitis had a 3.5 times higher odds to simultaneously have endometritis than animals without urocystitis.

Effects of prolonged exposure to low-dose fumonisin B1 in pigs.

From the point of view of human exposure, fumonisins (FB1, FB2, FB1, FB4), a relatively recently (1988) discovered and identified group of mycotoxins, represent one of the five most important mycotoxin groups causing human disease. In an earlier experiment studying the effects of relatively low doses (10, 20 and 40 p.p.m.) of FB1 in weaned piglets, it was established that the 4-week feeding of 10 p.p.m. (mg/kg feed) FB1 produced mild pulmonary oedema. This suggested the importance of studies with even lower doses of the toxin to determine the tolerable limits. The objective of this experiment was therefore to study the effects of prolonged (8-week) exposure to still lower concentrations (0, 1, 5 and 10 mg/kg feed) of FB1. The 8-week feeding of FB1 in low concentrations (1-10 p.p.m.) did not cause clinical signs and significant performance impairment in pigs, but rendered irreversible the chronic changes that had already developed in the animals in a dose-dependent manner. Dissection revealed pathological alterations of the lungs in one of the animals given 1 p.p.m. (n = 4), in two animals exposed to 5 p.p.m. (n = 5), and in three animals given 10 p.p.m. (n = 4). In all three treatment groups, proliferation of the connective tissue fibres, primarily of those around the lymphatic vessels, in the subpleural and interlobular connective tissue of the lungs, extending to the peribronchial and peribronchiolar areas, was seen. The results of this experiment call attention to the risk of prolonged low-dose toxin exposure, which has very important public health implications.

Preliminary communication: examination of the harmful effect to fetuses of fumonisin B(1) in pregnant sows.

Three sows were fed a diet mixed with Fusarium moniliforme fungal culture from the 107th day of pregnancy until parturition. Fumonisin B(1) toxin was administered to two sows (sows 1 and 2) in a daily dose of 300 mg for an additional 7 days subsequent to parturition, i.e., for a total of 14-16 days. The third sow (No. 3) was given the toxin in the same daily dose only until parturition, i.e., for 7 days in total. There were no symptoms observed in any of the sows. Two piglets were taken from each of the three sows and sacrificed immediately after parturition, i.e., prior to the first suckling. After 24 h, two additional piglets were taken for slaughter from each of the litters, which by then had access to colostrum. Finally, on the 7th day postparturition another two piglets per litter were sacrificed and material obtained from them was processed for examination. It was established that fumonisin B(1) present in the Fusarium moniliforme culture resulted in damage to the fetuses in utero. Of the changes indicating toxic effect, intraalveolar, subpleural, and interstitial pulmonary edema of various degrees of severity could be detected in the piglets sacrificed immediately following parturition and before the first suckling. Pathological changes were observed in the histopathological sections of the liver, and increases in the activities of plasma aspartic acid transaminase (AST), gamma glutamyl transferase (GGT) and alkaline phosphatase (AKLP), higher than physiological levels were detected. The serum-free sphinganine/sphingosine ratio, considered a bioindicator of fumonisin B(1) toxicosis, varied in accordance with the degree of severity of the changes which occurred. The values obtained were found to be between 0.29 and 0.36 in the cases of severe pulmonary edema, and between 0.20 and 0.24 for the cases of mild pulmonary edema. In the piglets of the sows fed the toxin for an additional 7 days subsequent to parturition and which were born with severe pulmonary edema, mild pulmonary edema could be detected after colostrum suckling, 24 h, and 7 days after parturition. The SA/SO value of the serum in these two piglets was 0.19 and 0.20, while at the same time AST, GGT, and ALKP values higher than physiological levels were measured. In the milk samples taken from sows 1 and 2 and examined after 24 h and after 7 days FB(1) was detected in quantities of 18.0-27.5 ppb. There were no changes observed on the seventh day in the piglets of the third sow, the diet of which contained no toxin after parturition. However, as the piglets of the third sow demonstrated only mild effects of pulmonary edema it is not possible to establish with certainty a postpartum cause-effect relationship between fumonisin in colostrum and pulmonary edema.

New human Dirofilarioses in Hungary.

About ten cases of filariosis have recently been recorded in the Hungarian medical literature, six of them caused by Dirofilaria repens. Dirofilaria repensis a mosquito-transmitted filaroid worm in the subcutaneous tissue of dogs and cats in the temperate areas of the Old World. It accidentally infects man, too, and can remain unidentified due to physicians, poor knowledge of the parasite. In the last two years six new Dirofilaria repensinfections have been found in various parts of the country: five localised dermally and one in the deep tissues. Two of the cases might have been acquired in Italy during summer travels. Four patients, however, have never been abroad, these cases must be considered autochtonous infections. The thickness of the multilayered cuticle of the worm, diameter of the body and the size, form and number of the longitudinal ridges on its surface are used in the histological diagnosis of the the parasite.

Experiment to determine limits of tolerance for fumonisin B1 in weaned piglets.

In Hungary almost 70% of mould-affected maize inspected since 1993 was found to be contaminated with fumonisin B1 (FB1) (mean 2.6-8.65 mg/kg; maximum 9.8-75.1 mg/kg), the degree of this contamination was found to increase from year to year (Fazekas et al., 1997b). In this experiment, in order to define tolerance limit values, the effect of exposing weaned piglets to FB1 in low doses over a 4-week period was examined. The experiment was performed with 20 weaned barrows of Danish Landrace breed. After a 5-day adaptation period cultures of the fungus Fusarium moniliforme were mixed into the animals' feed in concentrations that resulted in a daily intake of fumonisin B1 of 0, 10, 20 and 40 mg/kg feed. Feeding with the toxin was observed to exert no significant effect on body weight gain or feed consumption in the animals, no clinical signs were observed and no mortality traceable to toxic effects occurred. In computer tomography examinations performed in the second and fourth weeks mild and more severe pulmonary oedema was diagnosed in the experimental animals. The processes developing in the pulmonary parenchyma were corroborated by the mathematical and statistical evaluation procedures applied. The haematological parameters examined revealed no change attributable to toxic effects, while with respect to the biochemical parameters, an increase in aspartate aminotransferase (AST) activity dependent on dosage, indicating a pathological change in the liver, was ascertained in all three experimental groups. The free sphinganine to sphingosine ratio (SA/SO), which is regarded as the most sensitive bioindicator of fumonisin toxicosis, showed an increase proportionate to toxin concentration for all three dosages. Dissection revealed mild cases of pulmonary oedema in three of the animals given doses of 10 p.p.m. (n = 4), two mild and two severe cases in those exposed to 20 p.p.m. (n = 5), and severe cases in all five animals given 40 p.p.m. The oedema of non-inflammatory origin was confirmed by histopathological examinations. The findings of this experiment which indicate that in this study FB1 administered in substantially lower concentrations than those reported in the literature resulted in severe pathological changes, point to the importance of studies involving even lower doses.

Epitheliocystis disease in cultured pacu (Piaractus mesopotamicus) in Brazil.

Epitheliocystis disease was diagnosed in a cultured pacu (Piaractus mesopotamicus Holmberg, 1893) stock on the basis of gross pathological (greyish-white, pinpoint- or pinhead-sized focal areas, connected strongly to the gill lamellae) and histopathological examinations (hypertrophic cells 10-40 microns in size, having a well-defined wall and a central inclusion or characteristic granular content stained with Giemsa). The disease did not occur among fishes (Cyprinus carpio L., Ctenopharyngodon idella Cuv. and Val., Sarotherodon mosambicus Peters, Hoplias malabaricus Bloch) kept together with the affected pacus. About 30-40% of the pacu stock succumbed. Since branchial lesions and heavy mortality were observed in this pacu population, the aetiological role of the epitheliocystis agent could be suspected. Based upon ultrastructural examinations this agent is considered to be a chlamydia or a chlamydia-like organism.

Human Dirofilaria repens infection in Hungary: a case in the spermatic cord and a review of the literature.

Orchiectomy was performed in a 37-year-old Hungarian man exhibiting a swelling in his right testicle. Histology revealed a nodule attached to the spermatic cord, consisting of a granulomatous tissue around sections of a nematode. The worm was identified as Dirofilaria repens, an uncommon parasite in Hungary. As the patient had been abroad only in Italy where cases of dirofilariosis in dogs and humans are relatively frequent, it is assumed that the infection might have been acquired in that country 5 years earlier. This is the fifth case, published so far in the world, of such a localization in a human. The human cases of dirofilariosis reported in Hungary are reviewed.

Detection of Lawsonia intracellularis in Hungarian swine herds by polymerase chain reaction.

Pooled faecal samples and/or intestinal contents from pig carcasses were collected from 11 Hungarian swine farms and subjected to DNA extraction/ purification and subsequent polymerase chain reaction (PCR) in order to detect Lawsonia intracellularis, the aetiological agent of the porcine proliferative enteropathy complex. Specific PCR positivity was detected in 6 individual and 22 pooled samples out of 46, originating from eight herds. The PCR products of collected faecal samples hybridised in Southern blot hybridisation with the DNA of the type strain L. intracellularis NCTC 12657. This is the first confirmed detection of L. intracellularis by PCR in Hungary.

Paraintestinal mesenteric abscess and chronic peritonitis in a bull.

A 2-year-old bull was examined because of intermittent anorexia, signs of mild colic, and weight loss of 3 weeks' duration. A tympanitic resonance (ping) could be heard during simultaneous auscultation and percussion of the right paralumbar fossa, and a mass could be felt in the right dorsal quadrant of the abdominal cavity during palpation per rectum. Right flank laparotomy was performed, and intraoperative ultrasonography and ultrasound-guided fine-needle aspiration were used to determine that the mass was an abscess. However, the abscess could not be removed or drained into the colon because of extensive adhesions to other organs. Because the owner refused to pursue continued medical treatment, the bull was euthanatized. At necropsy, the abscess was found to be connected to a caudal mesenteric lymph node through a fistula. Histologic evaluation of the lymph node revealed hyperplastic lymphadenitis, and an alpha-hemolytic streptococcus was recovered from the abscess fluid. The most likely possibility for the findings in this bull were that the lymphadenitis was of hematogenous origin and that the abscess developed as a direct extension of the infectious process, similar to development of mesenteric abscesses in horses with chronic streptococcal infection (i.e., strangles).

Hepatic ultrasonographic findings in experimental carbon tetrachloride intoxication of the dog.

An experimental study was undertaken in dogs with carbon tetrachloride (CCl4) intoxication to describe and follow the changes in the ultrasound appearance of the liver. Characteristic ultrasound features (enlarged liver, increased echogenicity with fine, tightly-packed echoes, and loss of echogenicity of the portal vein walls) were seen from day 2 of the experiment, culminating between days 2 and 7. Subsequently the liver gradually and almost completely regained its normal ultrasound appearance. Ultrasonographic findings corresponded to the severity of the clinicopathological parameters, and changed parallel with the results of liver biopsies. Histology of the biopsy samples revealed acute centrolobular lipid accumulation with necrobiosis and necrosis in the hepatocytes in the first stage of the disease. In the second stage, a secondary reparatory-inflammatory process and reparation in the interstitial tissue were seen. In some dogs, ill-defined hyperechoic foci were observed within the liver during the regenerative phase. Hepatic ultrasonography seems to be a reliable and relatively sensitive method for monitoring liver dystrophy with necrobiosis and lipid accumulation and to follow the course of steatosis in clinical cases. Ultrasound-guided biopsy can further improve the diagnostic accuracy of ultrasonography in liver dystrophy.

Ultrasonographic findings of renal dysplasia in cocker spaniels: eight cases.

A retrospective study of eight young Cocker Spaniels aged 9-24 months was performed to describe the ultrasonographic findings of histologically confirmed renal dysplasia. Ultrasonography revealed kidneys of significantly (p < 0.001) reduced volume in all dogs. During qualitative evaluation, two different types of sonographic alterations could be seen. In one type of the ultrasound alterations, corticomedullary demarcation was distinct and the renal cortex was remarkably thin, which was best seen in the dorsal (frontal) imaging plane. In the other type of the ultrasound appearance, overall increased echogenicity with poor corticomedullary demarcation was noticed, and the kidneys could hardly be separated from their surroundings. These features were best recognised in the sagittal (coronal) imaging plane. In one dog with secondary hypercalcaemia, a hyperechoic corticomedullary area was also seen. Post-mortem histological diagnosis revealed renal dysplasia and secondary fibrosis. Based on ultrasound findings alone, renal dysplasia (renal familial disease) can be suspected when small kidneys with thin echogenic cortex are present in young dogs. An ultrasound image, similar to that of fibrotic kidneys (increased overall echogenicity and reduced corticomedullary definition) cannot be differentiated from chronic inflammatory disease and from end-stage kidneys. Therefore, ultrasound-guided biopsy or post-mortem histology is necessary for the definitive diagnosis of renal dysplasia. This is the first study reporting on the ultrasound appearance of renal dysplasia in Cocker Spaniel dogs.

The first isolation of Eubacterium suis in Hungary.

Eubacterium suis was isolated from the preputium of seven out of 16 mature boars on two farms and from the urinary bladder of one out of five sows originating from a third herd. The morphological and biochemical characteristics of the isolated strains were identical to that of the reference strain of E. suis ATTC 33144. Three out of four strains isolated from Farm A were successfully subcultured aerobically, and then anaerobically again. E. suis together with Proteus mirabilis was isolated from cystitis of a sow 4 days after artificial insemination. These are the first strains of E. suis isolated in Hungary.

Histological changes in the swimbladder wall of eels due to abnormal location of adults and second stage larvae of Anguillicola crassus.

In severe Anguillicola crassus infection of eels, adult helminths and 2nd stage larvae staying in the swimbladder lumen may occasionally get, through minor lesions of the tunica interna, into the subserosa of the swimbladder wall where they die and disintegrate. A thin connective tissue capsule is formed around the helminths that behave as foreign bodies in intercellular location, while the lacunas of the surrounding loose connective tissue comprise melanin-containing macrophages. In the environment of the 2nd stage larvae the formation of giant cells is a typical finding.

Complex study of the physiological role of cadmium. III. Cadmium loading trials on broiler chickens.

Cadmium (Cd) loading trials were conducted on a total of 110 (3 x 10 and 4 x 20) broiler chickens prereared for 21 days. The control chickens received no cadmium, while chickens in the six treatment groups were given different doses of Cd as an aqueous solution of CdSO4 administered either into the crop or mixed in the feed. The chickens were kept in a climatized animal house and treated usually for 3-5 weeks (maximum 68 days), with the exception of group Cd-75 chickens which were treated up to 239 days of age. The chickens' health status, body mass and feed consumption were monitored throughout the trial. On days 14-20 and on day 42 of the trial 2 chickens per group, then at the end of trial a total of 25 chickens were killed in anaesthesia. These birds, together with chickens that died or were killed during the trial, were subjected to detailed gross pathological examination. From 11 organs (kidney, liver, spleen, testicle, brain, myocardium, skeletal muscle, lungs, digestive tract, pancreas, tubular bones) of these chickens samples were taken for assay for a total of 16 elements, as well as for light and electron microscopic examination. With the exception of groups Cd-30 and Cd-600, no abnormal clinical signs were observed in the first two weeks of the trial. Chickens of group Cd-30 died before day 8-12 of the trial among signs of complete anorexia, rapid emaciation, huddling and diarrhoea, while chickens of group Cd-600 died before day 28, showing similar clinical signs. The body mass of chickens fed a Cd-supplemented diet either remained constant or decreased substantially, in a degree proportional to the Cd load. The only exception was group Cd-2.5, in which the average body mass of birds at the end of week 8 slightly exceeded that of the controls. Four out of the 10 cockerel chicks fed a diet containing 75 ppm Cd up to 239 days of age died of intercurrent diseases; the remaining six grew well and reached a body mass of 3.8-4.3 kg. Feed conversion efficiency was satisfactory in the control group and in group Cd-2.5 (2.1 and 2.4 kg, respectively) and could not be evaluated in a realistic manner in the other groups. At necropsy, the cockerel chicks of groups Cd-30 and Cd-600 showed severe emaciation, liver and kidney degeneration, myocardial hypertrophy and cardiac dilatation.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of long-term aluminum exposure on certain serum constituents in broiler chickens.

The aim of this work was to study the effects of 35 days exposure to aluminium on certain serum biochemical quantities in chickens. Broiler chicks (TETRA-726 hybrid, male) were kept in a climate-controlled stall with feed and water ad libitum, from day 1 of age, for 7 weeks. From the beginning of the third week aluminium was added to the diet as aluminium chloride. Treatments included supplemental aluminium content of 0, 200, 1000 and 3000 mg/kg ration. At the end of the experiment, blood samples were taken from the v. ulnaris. The treated groups showed significantly elevated alkaline phosphatase activities, as well as increased cholesterol concentrations and decreased triacylglycerol concentrations, and these changes were dose-dependent. The concentration of uric acid was significantly higher in the group receiving 1000 mg/kg ration, but significantly lower in the group receiving 3000 mg/kg ration, compared with the controls. In the treated groups, the concentration of glucose, as well as the activities of cholinesterase, aspartate aminotransferase, gamma-glutamyl transferase and creatine kinase were similar in the controls and treated animals. High levels of alkaline phosphatase are due to increased osteoblastic activity, provoked by the disturbance of bone formation, caused in turn by aluminium. Alterations in serum uric acid may be connected with metabolic disturbances (e.g. renal function, cation--anion balance etc.). Neither hepatic nor muscle damage was found.

Complex study of the physiological role of aluminium. II. Aluminium tolerance tests in broiler chickens.

Aluminium (Al) tolerance tests were carried out in 8 x 24 (a total of 192) broiler chickens pre-reared for 17 days. Chickens of the control group were fed a standard poultry grower diet ad libitum, while those in six experimental groups received a diet supplemented with different (200, 500 and 1,000 mg/kg of feed) doses of Al supplied in the form of AlCl3 and, in two cases, with the same dose of P added to the feed in the form of MCP, up to 52 days of age. Chickens of one group were fed a diet containing 3,000 mg Al per kg of feed for 70 days. The acidic pH of AlCl3 solution sprayed onto the feed was neutralized by adding a sufficient amount of NaHCO3 solution in all but one group. The birds' health status was monitored regularly throughout the feeding trial. Their body mass gain and feed consumption were recorded weekly. At the end of the trial, 3-10 chickens per group (a total of 40 birds) were exsanguinated, subjected to gross pathological examination, and samples were taken from 9 organs (liver, kidney, skeletal muscle, myocardium, brain, spleen, testicle, lungs and tubular bones) for light and electron microscopic examination and for the regular determination of 8 elements (Ca, P, Mg, Zn, Fe, Mn, Cu, Al). In addition, samples were taken from the organs of 2-5 chickens per group for analysis for additional 6 elements (Mo, Co, Ni, Sr, Na, K). From the results obtained the following main conclusions were drawn: (i) With the exception of four birds culled during the trial, all chickens remained symptomless throughout, and all chickens except those fed a diet supplemented with 3,000 mg/kg aluminium developed properly. (ii) The body mass of chickens in groups fed a diet supplemented exclusively with Al decreased moderately (by 67, 69 and 88 g, respectively), depending on Al concentration of the diet. Chickens fed a diet containing 3,000 mg/kg Al showed a very substantial (621.3 g; 32%) decrease in body mass Phosphorus supplementation did not markedly affect the body mass gain. (iii) Specific feed utilization was satisfactory (2.100-2.210 g/kg body mass). The very poor feed utilization of chickens subjected to the heaviest Al load can be attributed to the temporary disturbance of acid-base balance caused by the non-buffered AlCl3 solution.(ABSTRACT TRUNCATED AT 400 WORDS)

Comparative studies on susceptibility of the mouse (Mus musculus) and the vole (Microtus arvalis) to Listeria monocytogenes and Erysipelothrix rhusiopathiae.

The susceptibility of mice and voles to intraperitoneal and subcutan injection of virulent strains has been tested comparatively on 261 mice and 120 voles. Using various germ-doses, 50 per cent lethality has been established. Listeria monocytogenes serovar 1/2a killed mice in doses of 120,000 and 1.5 X 10(6) germs when injected intraperitoneally and subcutaneously and voles in doses of 1800 and 12,000 germs, respectively. Thus voles proved 100 to 120 times more susceptible than mice. Erysipelothrix rhusiopathiae killed mice injected both intraperitoneally or subcutaneously in minimal doses, such as 25 germs, but voles survived the infection with 1 to 5 million germs. This indicates that voles are resistant to E. rhusiopathiae, but more susceptible to L. monocytogenes than mice. In contrast, both these rodent species proved susceptible to the obligate pathogenic Francisella tularensis and invariably succumbed to septicaemia when injected with a few of these bacteria.