Comparing the hierarchy of inter- and intra-species interactions with population dynamics of wine yeast cocultures.

In winemaking, the development of new fermentation strategies, such as the use of mixed starter cultures with Saccharomyces cerevisiae (Sc) yeast and non-Saccharomyces (NS) species, requires a better understanding of how yeasts interact, especially at the beginning of fermentation. Despite the growing knowledge on interactions between Sc and NS, few data are available on the interactions between different species of NS. It is furthermore still unclear whether interactions are primarily driven by generic differences between yeast species or whether individual strains are the evolutionarily relevant unit for biotic interactions. This study aimed at acquiring knowledge of the relevance of species and strain in the population dynamics of cocultures between five yeast species: Hanseniaspora uvarum, Lachancea thermotolerans, Starmerella bacillaris, Torulaspora delbrueckii and Sc. We performed cocultures between 15 strains in synthetic grape must and monitored growth in microplates. Both positive and negative interactions were identified. Based on an interaction index, our results showed that the population dynamics seemed mainly driven by the two species involved. Strain level was more relevant in modulating the strength of the interactions. This study provides fundamental insights into the microbial dynamics in early fermentation and contribute to the understanding of more complex consortia encompassing multiple yeasts trains.

Mycobacterium leprae diversity and population dynamics in medieval Europe from novel ancient genomes.

BACKGROUND: Hansen's disease (leprosy), widespread in medieval Europe, is today mainly prevalent in tropical and subtropical regions with around 200,000 new cases reported annually. Despite its long history and appearance in historical records, its origins and past dissemination patterns are still widely unknown. Applying ancient DNA approaches to its major causative agent, Mycobacterium leprae, can significantly improve our understanding of the disease's complex history. Previous studies have identified a high genetic continuity of the pathogen over the last 1500 years and the existence of at least four M. leprae lineages in some parts of Europe since the Early Medieval period. RESULTS: Here, we reconstructed 19 ancient M. leprae genomes to further investigate M. leprae's genetic variation in Europe, with a dedicated focus on bacterial genomes from previously unstudied regions (Belarus, Iberia, Russia, Scotland), from multiple sites in a single region (Cambridgeshire, England), and from two Iberian leprosaria. Overall, our data confirm the existence of similar phylogeographic patterns across Europe, including high diversity in leprosaria. Further, we identified a new genotype in Belarus. By doubling the number of complete ancient M. leprae genomes, our results improve our knowledge of the past phylogeography of M. leprae and reveal a particularly high M. leprae diversity in European medieval leprosaria. CONCLUSIONS: Our findings allow us to detect similar patterns of strain diversity across Europe with branch 3 as the most common branch and the leprosaria as centers for high diversity. The higher resolution of our phylogeny tree also refined our understanding of the interspecies transfer between red squirrels and humans pointing to a late antique/early medieval transmission. Furthermore, with our new estimates on the past population diversity of M. leprae, we gained first insights into the disease's global history in relation to major historic events such as the Roman expansion or the beginning of the regular transatlantic long distance trade. In summary, our findings highlight how studying ancient M. leprae genomes worldwide improves our understanding of leprosy's global history and can contribute to current models of M. leprae's worldwide dissemination, including interspecies transmissions.

Spatiotemporal association between the mite Brevipalpus yothersi and Citrus leprosis virus C in orange orchards.

Citrus leprosis virus C (CiLV-C) is an economically important pathogen and the main causative agent of leprosis disease in citrus orchards. The main vector of this disease, the mite Brevipalpus yothersi, is widely distributed in Mexican orchards on a wide range of citrus species. Despite the importance of both the virus and the mite, field studies recording their occurrence and co-occurrence are practically non-existent. We systematically sampled orange orchards for both CiLV-C and B. yothersi throughout the year. The distribution of the CiLV-C and B. yothersi was evaluated on each sampling occasion and their spatiotemporal associations were determined. Specifically, 100-112 orange trees, distributed in 18 rows (five or six trees per row), were sampled monthly between March 2017 and February 2018 (11 sampling dates). Twenty leaves per tree were sampled on each occasion. The number of mites per tree and the percentage of leaves per tree with disease symptoms were recorded. On each sampling occasion, spatiotemporal associations between mites and disease were determined using the Spatial Analysis by Distance Indices (SADIE) method. CiLV-C and B. yothersi were identified using molecular methods. Throughout the study, the distribution of CiLV-C was aggregated and the distribution of B. yothersi was random. No association was found between the virus and the mite on any of the sampling dates. In total, 173 mites were collected, but only 43 mites were found to be carrying CiLV-C. The reason for this lack of association between the virus and the mite, as well as the impact of our findings on the epidemiology of the disease in orange orchards, are discussed.

Morphological and physiological assessments reveal that freshwater turtle (Mauremys leprosa) can flourish under extremely degraded-polluted conditions.

Freshwater turtles are long-lived sedentary organisms used as biological sentinels to assess anthropogenic perturbations in freshwater-ecosystems; notably because pollutants tend to accumulate in their tissues. Pollution has detrimental effects in sea turtles, but studies in freshwater turtles have provided contrasted results: several species have been impacted by habitat perturbation and pollution while others not. It is important to explore this issue since freshwater turtles are threatened worldwide. We compared two populations of the stripe necked terrapin (Mauremys leprosa) in a relatively pristine area (piedmont of the Atlas mountain) versus an extremely degraded-polluted area (sewers of a large city) in Morocco. All morphological and physiological proxies showed that turtles were able to cope remarkably well with highly degraded-polluted habitat. Population density, body size, and body condition were higher in the sewers, likely due to permanent water and food availability associated with human wastes. Stress markers (e.g. glucocorticoids) provided complex results likely reflecting the capacity of turtles to respond to various stressors. Reproductive parameters (testosterone level, indices of vitellogenesis) were lower in the relatively pristine area. The deceptive overall image provided by these analyses may hide the disastrous human impact on rivers. Indeed, Mauremys leprosa is the only aquatic vertebrate able to survive in the sewers, and thus, might nonetheless be a pertinent indicator of water quality, providing that the complexity of eco-physiological responses is considered.

Factors affecting prevailing population levels of Brevipalpus yothersi (Acari: Tenuipalpidae) in citrus areas affected by citrus leprosis in the State of Sao Paulo, Brazil.

Brevipalpus yothersi Baker (Acari: Tenuipalpidae) was recently confirmed as one of the main vectors of citrus leprosis. Knowledge about this mite's biology is essential to understand its population dynamics, in order to solve management issues in citrus orchards and explain why citrus leprosis is more severe in some regions. This paper aimed to study biological factors affecting prevailing population levels of B. yothersi and incidence of citrus leprosis. Mites were sampled from orchards in the south, north and northwestern regions of São Paulo State, Brazil. We assessed duration of the developmental stages, oviposition, incubation period, egg viability, longevity, net reproductive rate (Ro), mean generation time (T), intrinsic rate of increase (rm) and finite rate of increase (λ). There were small differences in parameter values between the three populations. Our results indicated that the various measures adopted in the control of the three populations did not lead to major biological differences between populations for the evaluated parameters.

Infestation dynamics of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) in citrus orchards as affected by edaphic and climatic variables.

Brevipalpus phoenicis (Geijskes) is a cosmopolitan and polyphagous mite that transmits important phytoviruses, such as coffee ringspot virus, passion fruit green spot virus and Citrus leprosis virus C. To characterise the dynamics of the probability and the rate of B. phoenicis infestation in response to edaphic and climatic factors, monthly inspections were performed in nine orchards in a citrus region of the State of Bahia, Brazil, for 35 months. Three fruits per plant were examined using a magnifying glass (10×) on 21 plants distributed along a "W"-shaped path in each orchard. Meteorological data were collected from a conventional station. To determine the correlations among the climatic variables, the data were analysed using Spearman correlations. Variables were selected by principal component analysis, and those that contributed the most to differentiate the groups were evaluated via a Mann-Whitney test. Using the quantile-quantile method, the limit values for the following climatic variables were determined: temperature (24.5 °C), photoperiod (12 h), relative humidity (83%), evapotranspiration (71 mm) and rainy days (14 days). The combination of longer days, higher temperatures, lower relative humidity levels and lower evapotranspiration increased the probability of B. phoenicis infestation, whereas successive rain events decreased that risk. Infestation rates were negatively affected by relative humidity levels above 83% and were positively affected by a decreasing available soil-water fraction and increasing insolation and photoperiod.

Yeast population dynamics during prefermentative cold soak of Cabernet Sauvignon and Malbec wines.

Prefermentative cold soak is a widely used technique in red wine production, but the impact on the development of native yeast species is hardly described. The aim of this work was to analyse the dynamics and diversity of yeast populations during prefermentative cold soak in red wines. Three different temperatures (14 ± 1 °C; 8 ± 1 °C and 2.5 ± 1 °C) were used for prefermentative cold soak in Cabernet Sauvignon and Malbec grape musts. Saccharomyces and non-Saccharomyces populations during cold soak and alcoholic fermentation were analysed. In addition, the impact on chemical and sensory properties of the wines was examined. Yeast dynamics during prefermentative cold soak were temperature dependent. At 14 ± 1 °C, the total yeast population progressively increased throughout the cold soak period. Conversely, at 2.5 ± 1 °C, the yeast populations maintained stable during the same period. Prefermentative cold soak conducted at 14±1°C favoured development of Hanseniospora uvarum and Candida zemplinina, whereas cold soak conducted at 8 ± 1 °C favoured growth of Saccharomyces cerevisiae. At 2.5 ± 1 °C, no changes in yeast species were recorded. Acidity and bitterness, two sensory descriptors, appear to be related to wines produced with prefermentative cold soak carried out at 14 ± 1 °C. This fact could be associated with the increase in non-Saccharomyces during the prefermentation stage. Our results emphasise the importance of the temperature as a determinant factor to allow an increase in non-Saccharomyces population during prefermentative cold soak and consequently to modify sensorial attributes of wines as well as their sensorial impact.

New record of Akashiwo sanguinea (Dinophyta) in the tropical estuarine waters of Northeastern Brazil (Western Atlantic).

This study reports the occurrence and the effect of the environmental factors on the spatial and temporal distribution of the dinoflagellate Akashiwo sanguinea (Hirasaka) Hansen & Moestrup in estuarine waters of northeastern Brazil. Samples were collected at seven stations from March 2007 to February 2008 during high tide and low tide, using Van Dorn bottles. The samples were immediately fixed with Lugol and analyzed with the Utermöhl method. Water samples were also collected for the identification of the hydrological characteristics of the area. Akashiwo sanguinea occurred throughout the annual cycle and at all sampling sites with densities ranging between 5 and 410 x 103 cells.L-1. The highest densities were recorded at low tide, especially during the months of the rainy season (July: 210 x 103 cells.L-1; August: 410 x 103 cells.L-1). Density values were within the normal range and blooms were not detected. Despite being common in the area, the species showed preference for sites with high concentrations of orthophosphate and total dissolved phosphorus and with salinity in the mesohaline regime.

New record of Akashiwo sanguinea (Dinophyta) in the tropical estuarine waters of Northeastern Brazil (Western Atlantic) / Novo registro de Akashiwo sanguinea (Dinophyta) em águas estuarinas tropicais do nordeste do Brasil (Atlântico Ocidental)

This study reports the occurrence and the effect of the environmental factors on the spatial and temporal distribution of the dinoflagellate Akashiwo sanguinea (Hirasaka) Hansen & Moestrup in estuarine waters of northeastern Brazil. Samples were collected at seven stations from March 2007 to February 2008 during high tide and low tide, using Van Dorn bottles. The samples were immediately fixed with Lugol and analyzed with the Utermöhl method. Water samples were also collected for the identification of the hydrological characteristics of the area. Akashiwo sanguinea occurred throughout the annual cycle and at all sampling sites with densities ranging between 5 and 410 x 103 cells.L–1. The highest densities were recorded at low tide, especially during the months of the rainy season (July: 210 x 103 cells.L–1; August: 410 x 103 cells.L–1). Density values were within the normal range and blooms were not detected. Despite being common in the area, the species showed preference for sites with high concentrations of orthophosphate and total dissolved phosphorus and with salinity in the mesohaline regime.

Este estudo relata a ocorrência e o efeito dos fatores ambientais na distribuição espacial e temporal do dinoflagelado Akashiwo sanguinea (Hirasaka) Hansen et Moestrup em águas estuarinas do Nordeste do Brasil. As amostras foram coletadas em sete estações, durante o período de março 2007 a fevereiro 2008 durante os regimes de preamar e baixa-mar, com o auxílio de garrafas tipo Van Dorn. As amostras foram imediatamente fixadas com lugol e analisadas pelo método de Utermöhl. Amostras de água foram também coletadas para a identificação das características hidrológicas da área. Akashiwo sanguinea ocorreu em todo o ciclo anual e em todos os locais de coleta com densidade variando de 5 a 410 cells.L–1 x 103, sendo as maiores densidades registradas na baixa-mar, principalmente, durante os meses do período chuvoso (julho 210 cells.L–1 x 103, agosto 410 cells.L–1 x 103). Os valores quantitativos estiveram dentro dos padrões normais, não tendo sido detectados florescimentos com conotações de blooms. Apesar de ser frequente na área, a espécie mostrou preferência pelos locais com maiores concentrações de ortofosfato dissolvido e fósforo total dissolvido e salinidade compreendida no regime mesoalino.

Motives and determinants for residence change after leprosy diagnosis, central Brazil.

OBJECTIVE: To determine the extent of population movement after diagnosis with leprosy and to describe the underlying motives and determinants for relocation. DESIGN: A cross-sectional study was conducted among those newly diagnosed with leprosy in 79 endemic municipalities in the state of Tocantins, central Brazil. Individuals were identified through the National Information System for Notifiable Diseases (SINAN) database and interviewed with structured questionnaires. RESULTS: In total, 224 (20.9%) out of 1070 individuals relocated after their diagnosis with leprosy. Respondents moved to another neighbourhood in the same municipality (n = 178, 79.5%), followed by another municipality in Tocantins state (n = 26, 11.6%) and in another state (n = 11, 4.9%). The primary motives and/or determinants for relocation were: home ownership (n = 55, 28.4%), familial reasons (n = 43, 19.2%), to seek better living conditions (n = 27, 13.9%), employment (n = 26, 11.6%), and better neighbourhood (n = 22, 9.8%). Other motives were related to better access to leprosy diagnosis/treatment (n = 11, 4.9%), owner-terminated rental (n = 5, 2.2%), personal finances/could not afford housing (n = 4, 1.8%). Perceived stigma due to leprosy was mentioned by one participant (0.5%). CONCLUSION: In Tocantins state, population movement is lower among individuals recently diagnosed with leprosy, as compared to the overall population. The primary motives for relocation after leprosy diagnosis were related to lifestyle changes. Stigma and treatment-related reasons did not appear to be common motives for population movement. These results may reflect policy changes instituted from the Brazilian Program of Leprosy Control to decentralise leprosy services and intensify health education campaigns within a broader concept of Information, Education and Communication.

Yeast population dynamics during pilot-scale storage of grape marcs for the production of Grappa, a traditional Italian alcoholic beverage.

The composition and population dynamics of the yeast microflora of grape marcs were investigated during a pilot scale fermentation study using two white grape varieties, namely Moscato and Prosecco, from two distinct areas of the Veneto Region. Yeast counts were made at the beginning, after 4 and after 15 days of marc storage under anaerobic conditions. Seventy isolates from each sampling time were identified to species by RAPD-PCR analysis and subsequent ITS region sequencing. A good biodiversity of yeasts occurred in both marcs at the beginning of fermentation, with high presence of Hanseniaspora opuntiae, but without detectable presence of Saccharomyces strains, which instead became the dominant yeast after just 4 days of fermentation, remaining at that level until the end of fermentation. Colonization of Moscato marc by S. cerevisiae resulted better, in relation to its higher sugar content. Characterization of S. cerevisiae isolates by mitochondrial DNA restriction analysis revealed the presence of 66 different strains in the marc from the Moscato grapes, without the occurrence of a clearly dominant strain, while in the marc from the Prosecco grapes only 23 different profiles were scored, with a dominant strain that accounted for 62.7% of the Saccharomyces population after 4 days of fermentation.

Genotypic analysis of Mycobacterium leprae strains from different regions of India on the basis of rpoT.

Mycobacterium leprae strains from Indian leprosy patients were analyzed using the six base tandem repeat, GACATC, in rpoT gene as genetic marker. DNA was extracted from slit-skin smears and nasal swabs of new untreated as well as treated leprosy patients living in different regions of India. PCR amplification of rpoT gene and sequencing of amplicons showed the presence of two genotype of M. leprae in this study, 73.4% having three copies (ancient Indian type) and 26.6% contain 4 copies (considered to be Japanese and Korean). These genotypes along with other short tandem repeats may help in studying the historical spread of disease and the strains of M. leprae disseminated by various human races that migrated to India from other places of Asia and European countries during our history.

Effect of oenological practices on microbial populations using culture-independent techniques.

Sulphur dioxide (SO(2)) addition and yeast inoculation are well-established practices in winemaking for restricting the growth of indigenous yeasts and bacterial populations. The effect of these oenological practices on wine microbial populations has been evaluated using culture-independent methods. These are quantitative PCR (qPCR) for the enumeration of yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB), and PCR-DGGE to determine the yeast and bacteria species diversity. The PCR-DGGE method detected a low yeast and bacteria species diversity. On the contrary, the specificity of the primers designed for the qPCR allowed that minor microbial groups such as Hanseniaspora were accurately quantified regardless of a large presence of other microbial groups such as Saccharomyces. From an oenological point of view, inoculation increased the proportion of Saccharomyces vs. non-Saccharomyces in a shorter time. Hanseniaspora increased during the first phase and decreased during the latter phases of the process, especially in the sulphited fermentations. Both yeast inoculation and SO(2) kept the LAB populations at very low level, while the AAB populations were hardly affected by these two practices.

Dynamics and diversity of non-Saccharomyces yeasts during the early stages in winemaking.

This detailed study observed the yeasts present in the ecological niche of "wine must". The dynamics and identity of non-Saccharomyces yeasts during the cold maceration and alcoholic fermentation of grape must were investigated under real production conditions in the Bordeaux region. Furthermore, we studied the impact of two oenological parameters on the development and diversity of non-Saccharomyces yeasts during cold maceration: temperature management and the timing of dried yeast addition. The non-Saccharomyces community underwent constant changes throughout cold maceration and alcoholic fermentation. The highly diverse non-Saccharomyces microflora was present at 10(4)-10(5) CFU/mL during cold maceration. The population increased to a maximum of 10(6)-10(7) CFU/mL at the beginning of alcoholic fermentation, then declined again at the end. The population at this point, evaluated at around 10(3)-10(4) CFU/mL, was shown to be dependent on the timing of yeast inoculation. The choice of temperature was the key factor for controlling the total yeast population growth, as well as the species present at the end of cold maceration. Hanseniaspora uvarum was a major species present in 2005 and 2006, while Candida zemplinina was very abundant in 2006. A total of 19 species were isolated.

Application of fluorescence in situ hybridisation (FISH) to the analysis of yeast population dynamics in winery and laboratory grape must fermentations.

To analyse the yeast population diversity during wine fermentations, specific fluorescein-labelled oligonucleotide probes targeted to the D1/D2 region of the 26S rRNA of different yeast species known to occur frequently in this environment were designed and tested with reference strains. The probes were then used to identify wine must isolates and to follow, in combination with plate counts, the evolution of yeast populations in two winery fermentations of white and red grape musts. In both cases, a high diversity of non-Saccharomyces yeast species was detected, including Candida stellata, Hanseniaspora uvarum, H. guilliermondii, Kluyveromyces marxianus, K. thermotolerans and Torulaspora delbrueckii. Some of these species (e.g., K. marxianus, K. thermotolerans and T. delbrueckii) were present in significant amounts during the tumultuous fermentation stage, despite the predominance of Saccharomyces cerevisiae cells following the inoculation of the wine musts with a starter strain. To further clarify the yeast population dynamics at the late phase of the fermentations, and because winery conditions do not allow a reliable control of experimental variables, strains isolated from the industrial musts were used to conduct two laboratory microvinifications in synthetic grape juice, using different ratios of S. cerevisiae/non-Saccharomyces in the inocula. Under these conditions, the results were similar to those obtained in the winery, showing a yeast profile with mixed species throughout the first fermentation stage, i.e. until about 40-50% of the total sugar was consumed. Non-Saccharomyces yeasts were outgrown by S. cerevisiae only after ethanol reached concentrations around 4-5% (v/v), which argues in favour of a potential important role of non-Saccharomyces in the final organoleptic characteristics of the wine.

Alcohols, esters and heavy sulphur compounds production by pure and mixed cultures of apiculate wine yeasts.

Strains of Hanseniaspora uvarum, Hanseniaspora guilliermondii and Saccharomyces cerevisiae were used as pure or mixed starter cultures in commercial medium, in order to compare their kinetic parameters and fermentation patterns. In pure and mixed cultures, yeasts presented similar ethanol yield and productivity. Pure cultures of H. uvarum and S. cerevisiae showed a specific growth rate of 0.38 h(-1); however, this value decreased when these yeasts were grown in mixed cultures with H. guilliermondii. The specific growth rate of pure cultures of H. guilliermondii was 0.41 h(-1) and was not affected by growth of other yeasts. H. guilliermondii was found to be the best producer of 2-phenylethyl acetate and 2-phenylethanol in both pure and mixed cultures. In pure cultures, H. uvarum led to the highest contents of heavy sulphur compounds, but H. guilliermondii and S. cerevisiae produced similar levels of methionol and 2-methyltetrahydrothiophen-3-one. Growth of apiculate yeasts in mixed cultures with S. cerevisiae led to amounts of 3-methylthiopropionic acid, acetic acid-3-(methylthio)propyl ester and 2-methyltetrahydrothiophen-3-one similar to those obtained in a pure culture of S. cerevisiae; however, growth of apiculate yeasts increased methionol contents of fermented media.

On the origin of leprosy.

Leprosy, a chronic human disease with potentially debilitating neurological consequences, results from infection with Mycobacterium leprae. This unculturable pathogen has undergone extensive reductive evolution, with half of its genome now occupied by pseudogenes. Using comparative genomics, we demonstrated that all extant cases of leprosy are attributable to a single clone whose dissemination worldwide can be retraced from analysis of very rare single-nucleotide polymorphisms. The disease seems to have originated in Eastern Africa or the Near East and spread with successive human migrations. Europeans or North Africans introduced leprosy into West Africa and the Americas within the past 500 years.

Diseases in European historical populations and their effects on individuals and society.

Traditionally, populations are considered "historical" when first written evidences appear. Cultural development with all it's features--agricultural innovations and as a consequence--population growth, social specialisation and stratification, and finally--urbanisation trigger new environmental conditions, among them also in epidemiological situation, that have the feedback effects on the population and the individual. At this time only qualitative evaluations on many of questions can be presented. This paper is an attempt to synthesize available data on the following specific topics related to Medieval/Early Modern European population welfare: nutritional intake, malnutrition and famine; great epidemics and their consequences; spreading of specific infectious diseases (tuberculosis, syphilis, leprosy), which also had a great social resonance in those times, professional diseases. Considerations on their impact on demographical situation (life expectancy, birth rate), life quality and physical development and body build of an individual are also presented. Finally, possibilities for directional selection and impact on modern genetic diversity in Europe are discussed.

Infectious diseases in the 21st century.

Infecto-contagious diseases in the twenty-first century with respect to precedent will see themselves deprived of smallpox, dracunculiasis and very probably of paralyzing poliomyelitis. Vaccination-preventable diseases, such as measles, whooping cough, diphtheria, tetanus, rabies, some forms of meningitis, yellow fever and episodes of disseminated tuberculosis will greatly diminish in their rates of morbi-lethality; the elimination of some, and the eradication of measles, are expected. Other diseases such as diarrhea (including cholera), geo-helminthiasis, some severe respiratory tract infections and the majority of vector-transmitted infectious diseases will decrease due to improvements in potable water services, drainage, sanitary food control, living quarters, and individual and community anti-vector action. Leprosy, onchocerciasis and several parasitoses will be controlled by the available antimicrobial drugs. Infectious diseases will continue to be an important health problem due to: Reduction in the immunocompetence resulting from the aging of the population, chemotherapies necessary for neoplasms, and autoimmune pathology and the survival of persons with primary immunodeficiencies; lifestyles prone to infectious pathology, such as mega-city urbanization, children in day care centers, industrialized foods, intravenous drug addiction, sexual liberation, global commerce, and tourism; antibiotic-multiresistant microbial flora; environmental disturbances as a result of global warming, deforestation, the settling of virgin areas, dams, the large-scale use of pesticides, fertilizers and antimicrobials, and natural/social disasters generators of poverty, violence and deprivation will result in emergence or re-emergence of infectious diseases already controlled in the past.

The pattern of 'drop-out' of smear-positive cases at an urban leprosy centre.

One hundred nineteen smear-positive leprosy cases registered at an urban leprosy centre in Bombay in 1991 were followed for three years to study the 'drop-out' pattern in them and judge the utility of some corrective measures for the same. The measures included having maps showing exact location of the patient's residence, paying home visits on registration days and subsequent persuasion and counselling both at the clinic and at the residence of patients. The results were compared with 'drop-out' in smear-positive cases registered at the same centre in 1989, 1990, 1992 and 1993. By introduction of the special measures, the 'drop-out' rate was significantly reduced from 52% (for other years) to 36% (1991). The expenses incurred for the successful recovery of 'drop-out prone' patients and ensuring regularity in drug intake was Rs. 659/- per patient. This study of 'drop-out' patient shows that there are three categories of the so-called drop-outs: (i) the false 'drop-outs' (51%): these patients get transfer as per their convenience to other leprosy centres or medical services (private practitioners or consultants) within the city (ii) drop-outs due to migration: the migration is forced on them due to some genuine reason, and (iii) persistent offenders: this is a group of adamant, non co-operative, or, distressed patients. For the first two category of patients it is advisable to introduce a good referral system. For the recalcitrant defaulters, supervised short-term drug therapy will probably be the best option.