Screening Method to Evaluate Amino Acid-Decarboxylase Activity of Bacteria Present in Spanish Artisanal Ripened Cheeses.

A qualitative microplate screening method, using both low nitrogen (LND) and low glucose (LGD) decarboxylase broths, was used to evaluate the biogenic amine (BA) forming capacity of bacteria present in two types of Spanish ripened cheeses, some of them treated by high hydrostatic pressure. BA formation in decarboxylase broths was later confirmed by High Performance Liquid Chromatography (HPLC). An optimal cut off between 10⁻25 mg/L with a sensitivity of 84% and a specificity of 92% was obtained when detecting putrescine (PU), tyramine (TY) and cadaverine (CA) formation capability, although these broths showed less capacity detecting histamine forming bacteria. TY forming bacteria were the most frequent among the isolated BA forming strains showing a strong production capability (exceeding 100 mg/L), followed by CA and PU formers. Lactococcus, Lactobacillus, Enterococcus and Leuconostoc groups were found as the main TY producers, and some strains were also able to produce diamines at a level above 100 mg/L, and probably ruled the BA formation during ripening. Enterobacteriaceae and Staphylococcus spp., as well as some Bacillus spp. were also identified among the BA forming bacteria isolated.

High pressure processing effect on different Listeria spp. in a commercial starter-free fresh cheese.

In this study, both microbial inactivation and growth of Listeria spp. inoculated in commercial free-starter fresh cheese was evaluated after high-pressure processing (HPP). HPP conditions (300, 400, 500 and 600 MPa at 6 °C for 5 min) and inoculum level (3-4 or 6-7 log CFU/g of cheese), as well as differences among strains inoculated (Listeria innocua, L. monocytogenes CECT 4031 and L. monocytogenes Scott A) were investigated. Inactivation and generation of sublethal injury were determined after HPP using ALOA (Agar Listeria according to Ottaviani and Agosti) and TAL (Thin Agar Layer) plating methods, respectively. Listeria inactivation increased with the pressure applied, presenting some statistical differences between the employed strains, inoculum level and sublethal injury. The highest lethality values were obtained at 600 MPa for the three strains tested, although the 500 MPa treatment presented high lethality for L. innocua and L. monocytogenes CECT 4031. After treatment, L. innocua and L. monocytogenes CECT 4031 counts in fresh cheese increased gradually during cold storage. By contrast, counts in cheeses inoculated with L. monocytogenes Scott A did not change significantly (p ≥ 0.05), being this strain the most pressure resistant and with the slowest growth rate. The manuscript present information supporting that, strains with high-level resistance should be employed during inactivation studies, instead of surrogate microorganisms. Application of HPP treatments of 500 MPa and especially 600 MPa on fresh cheeses would be effective to eliminate the most resistant microorganism to a level that should not present a public health risk under normal conditions of distribution and storage.

Mordedura por víbora de Seoane. Descripción de un caso y revisión de la literatura / Vipera seoanei bite. Report of a case and review of the literature

No disponible

Inactivation of Bacillus spores inoculated in milk by Ultra High Pressure Homogenization.

Ultra High-Pressure Homogenization treatments at 300 MPa with inlet temperatures (Ti) of 55, 65, 75 and 85 °C were applied to commercial Ultra High Temperature treated whole milk inoculated with Bacillus cereus, Bacillus licheniformis, Bacillus sporothermodurans, Bacillus coagulans, Geobacillus stearothermophilus and Bacillus subtilis spores in order to evaluate the inactivation level achieved. Ultra High-Pressure Homogenization conditions at 300 MPa with Ti = 75 and 85 °C were capable of a spore inactivation of ∼5 log CFU/mL. Furthermore, under these processing conditions, commercial sterility (evaluated as the complete inactivation of the inoculated spores) was obtained in milk, with the exception of G. stearothermophilus and B. subtilis treated at 300 MPa with Ti = 75 °C. The results showed that G. stearothermophilus and B. subtilis have higher resistance to the Ultra High-Pressure Homogenization treatments applied than the other microorganisms inoculated and that a treatment performed at 300 MPa with Ti = 85 °C was necessary to completely inactivate these microorganisms at the spore level inoculated (∼1 × 10(6) CFU/mL). Besides, a change in the resistance of B. licheniformis, B. sporothermodurans, G. stearothermophilus and B. subtilis spores was observed as the inactivation obtained increased remarkably in treatments performed with Ti between 65 and 75 °C. This study provides important evidence of the suitability of UHPH technology for the inactivation of spores in high numbers, leading to the possibility of obtaining commercially sterile milk.

Concentración del fragmento aminoterminal del NT-proBNPen plasma: ¿un nuevo marcador biológico predictivo en las neumonías adquiridas en la comunidad? / N-terminal fragment of pro-brain natriuretic peptide plasma concentration: a new predictive biomarker for community acquired pneumonia?

Objetivo: Evaluar la relación entre la concentración plasmática del fragmento aminoterminal del pro-péptido natriurético cerebral (NT-ProBNP) en el momento del diagnóstico de neumonía adquiridad en la comunidad (NAC) en urgencias y su gravedad determinada como mortalidad a los 30 días. Método: Estudio observacional prospectivo tipo cohortes en el que se determina NTProBNP (medido en ng/l) como factor exposición, se realiza seguimiento durante 30 días y se analiza la frecuencia de mortalidad. Incluimos a los pacientes con diagnóstico clínico-radiográfico de NAC atendidos consecutivamente en el servicio de urgencias desde el 1 de febrero al 30 de abril de 2012. Resultados: Se valoraron 110 pacientes, y 96 fueron incluidos en el estudio (87%), 44% mujeres. La edad media fue 66 (DE 22) años. Los valores medios en sangre de NT-Pro BNP extraídos fueron de 3.747 ng/l (DE 10.732) y mediana 506 ng/l (RIC2.490). Fallecieron 13 pacientes en los primeros 30 días (14%). Los valores de NTproBNP medios en los pacientes fallecidos fueron 17.805 ng/l (DE 24.952) comparados con 1.545 ng/l (DE 2.467) en los pacientes supervivientes (p = 0,003). Conclusiones: Los valores de NT-ProBNP en el momento del diagnóstico de NAC en elSU son un buen predictor de mortalidad precoz a los 30 días (AU)

Objective: To assess the association between plasma concentration of the N-terminal fragment of the precursor to brain type natriuretic peptide (NT-proBNP) and severity of community-acquired pneumonia (CAP) (30-day mortality) at the moment of emergency department diagnosis. Methods: Prospective observational cohort study of NT-proBNP plasma concentration as the potential indicator of risk of 30-day mortality. Emergency department patients with a clinical-radiographic diagnosis of CAP were enrolled consecutively from February 1 to April 30, 2012. Results: A total of 110 patients were recruited; 96 (87%) were included. Women comprised 44% of the cohort. The mean (SD) age was 66 (22) years. The mean NT-proBNP concentration was 3747 (10 732) ng/L; the median concentration was 506 ng/L (interquartile range, 2490 ng/L). Thirteen patients (14%) died within 30 days. The mean NT-proBNP concentration was 17 804.85 (24 952) ng/L in patients who died and 1545 (2467) ng/L in survivors (P=.003). Conclusions: NT-proBNP concentration at the time of CAP diagnosis in the emergency department provides a good predictor of early (30-day) mortality (AU)

Charcot neuroarthropathy triggered and complicated by osteomyelitis. How limb salvage can be achieved.

BACKGROUND: Charcot neuroarthropathy is a severe complication in the feet of patients with diabetes, which can lead to a major amputation. Osteomyelitis and surgery for osteomyelitis have been reported as trigger mechanisms of developing Charcot neuroarthropathy. However, the development of acute Charcot neuroarthropathy triggered by osteomyelitis during conservative antibiotic treatment is not well outlined in the medical literature. CASE REPORTS: Two patients apparently developed mid and rear foot Charcot neuroarthropathy, which was clinically suspected while being treated with antibiotics for osteomyelitis. One of them presented osteomyelitis of the navicular bone and subsequently developed acute Charcot neuroarthropathy of the tarsometatarsal joints. The other presented calcaneal osteomyelitis with pathological fracture and developed Charcot neuroarthropathy of the transverse tarsal joint. No offloading had been implemented in either case. A major amputation had been indicated in both cases in their teaching hospitals. Limb salvage was achieved in both cases by means of surgery, culture-guided post-operative antibiotics, intraosseus instillation of super-oxidized solution, bed rest before placing a total contact cast and stabilization of the unstable foot with a total contact cast with an opening for checking the healing course and to detect any complications. The mechanisms of the development of acute Charcot neuroarthropathy in a patient with osteomyelitis are discussed. CONCLUSIONS: Osteomyelitis in the feet of patients with diabetes and neuropathy may trigger the development of acute Charcot neuroarthropathy. Fractures and dislocated joints may subsequently become infected from the index focus, producing a severe infected and unstable foot that may require a major amputation. Limb salvage can be achieved in specialized departments.

Does osteomyelitis in the feet of patients with diabetes really recur after surgical treatment? Natural history of a surgical series.

AIMS/HYPOTHESIS: The aim of this study was to determine the rate of recurrence, reulceration and new episodes of osteomyelitis and the duration of postoperative antibiotic treatment in a prospective cohort of patients with diabetes who underwent conservative surgery for osteomyelitis. METHODS: The prospective cohort included patients with diabetes and a definitive diagnosis of osteomyelitis who were admitted to the Diabetic Foot Unit (Surgery Department, La Paloma Hospital, Las Palmas de Gran Canaria, Spain) and underwent surgical treatment from 1 November 2007 to 30 May 2010. RESULTS: Eighty-one patients were operated on for osteomyelitis during the study period. Seven patients were lost to follow-up at different stages of the study. The median duration of follow-up was 101.8 weeks (quartile 1 = 56.6, quartile 3 = 126.7). Forty-eight patients (59.3%) underwent conservative surgery, 32 (39.5%) had minor amputations and there was one (1.2%) major amputation. Twenty patients (24.7%) required reoperation because of persistent infection. Postoperative antibiotic treatment over a median period of 36 days was provided. Wound healing was achieved by secondary intention for a median of 8 weeks. Sixty-five patients were available for follow-up after healing. The percentage of recurrence, reulceration, and new episodes of osteomyelitis was 4.6% (3/65), 43% (28/65) and 16.9% (11/65), respectively. Mortality during follow-up (excluding in-hospital deaths and patients lost to follow-up) was 13% (9/69). CONCLUSION: A low rate of recurrence of osteomyelitis after surgical treatment for osteomyelitis was achieved. Despite new episodes, our approach to managing this cohort of patients with diabetes and foot osteomyelitis achieved 98.8% limb salvage.

Triggering mechanisms of neuroarthropathy following conservative surgery for osteomyelitis.

BACKGROUND: The purpose of this study was to raise awareness and stimulate discussion of the possible triggering factors of Charcot neuroarthropathy by presenting the case of one patient who had both undergone surgery and was suffering from osteomyelitis. CASE REPORT: We have extracted one case from our data set for a patient who underwent conservative surgery for osteomyelitis and subsequently developed acute Charcot in the midfoot. We present the clinical findings, photographs and X-ray studies. Preoperative X-ray showed irregular severe bone destruction in the fourth metatarsal head and a fracture of the fourth metatarsal bone. No signs of midfoot Charcot neuroarthropathy were found in this preoperative X-ray. The third and fourth metatarsal bones were both removed and the surgical wound was left open to heal by second intention. Histopathological study confirmed osteomyelitis in the bone sample. Twenty-five days after surgery, the surgical wound showed no signs of infection and healing progressed in a satisfactory way. However, the foot was swollen, erythematous and warm. Skin temperature was two degrees higher than the contralateral foot. X-ray was taken and acute neuroarthropathy of the tarso-metatarsal joints was diagnosed. CONCLUSIONS: Charcot neuroarthropathy appears to have been triggered by bone infection and/or surgery. We believe that the pivotal factor in the development of acute Charcot neuroarthropathy in this case was the weight bearing in the deformed foot so soon after the operation. Immobilization of the foot is critical as it serves to decrease the inflammation which has a key role in the development of Charcot neuroarthropathy.

Fat content increases the lethality of ultra-high-pressure homogenization on Listeria monocytogenes in milk.

Listeria monocytogenes CCUG 15526 was inoculated at a concentration of approximately 7.0 log(10) cfu/mL in milk samples with 0.3, 3.6, 10, and 15% fat contents. Milk samples with 0.3 and 3.6% fat content were also inoculated with a lower load of approximately 3.0 log(10) cfu/mL. Inoculated milk samples were subjected to a single cycle of ultra-high-pressure homogenization (UHPH) treatment at 200, 300, and 400 MPa. Microbiological analyses were performed 2 h after the UHPH treatments and after 5, 8, and 15 d of storage at 4 degrees C. Maximum lethality values were observed in samples treated at 400 MPa with 15 and 10% fat (7.95 and 7.46 log(10) cfu/mL), respectively. However, in skimmed and 3.6% fat milk samples, complete inactivation was not achieved and, during the subsequent 15 d of storage at 4 degrees C, L. monocytogenes was able to recover and replicate until achieving initial counts. In milk samples with 10 and 15% fat, L. monocytogenes recovered to the level of initial counts only in the milk samples treated at 200 MPa but not in the milk samples treated at 300 and 400 MPa. When the load of L. monocytogenes was approximately 3.0 log(10) cfu/mL in milk samples with 0.3 and 3.6% fat, complete inactivation was not achieved and L. monocytogenes was able to recover and grow during the subsequent cold storage. Fat content increased the maximum temperature reached during UHPH treatment; this could have contributed to the lethal effect achieved, but the amount of fat of the milk had a stronger effect than the temperature on obtaining a higher death rate of L. monocytogenes.

Are diabetic foot ulcers complicated by MRSA osteomyelitis associated with worse prognosis? Outcomes of a surgical series.

AIMS: The aim of this study was to compare the outcomes of surgical treatment of osteomyelitis caused by methicillin-resistant Staphylococcus aureus (MRSA) with cases caused by methicillin-sensitive Staphylococcus aureus (MSSA). METHODS: We abstracted data of a series of 185 consecutive patients with diabetes and foot osteomyelitis undergoing surgery within the first 12 h after admission at a single hospital. Bone infection was confirmed by histopathological studies. Only cases where Staphylococcus aureus was isolated from bone specimens were included in this analysis. We analysed several variables between the two groups: MRSA vs. MSSA. RESULTS: MRSA bone infection was associated with higher body temperature (P = 0.02) and white blood cell count (P = 0.02) than MSSA. Patients with MRSA infections underwent a greater number of surgical procedures (P = 0.03). Limb salvage was achieved in 93.6% of the patients, with no statistically significant difference in limb salvage rates between MRSA and MSSA-related osteomyelitis. CONCLUSIONS: From our experience, where treatment is based on early and aggressive surgical treatment, MRSA bone infections are not associated with worse prognosis.

Inactivation of Salmonella enterica serovar Senftenberg 775W in liquid whole egg by ultrahigh pressure homogenization.

Two batches of samples of liquid whole egg were inoculated with a load of approximately 3 and 7 log CFU/ml, respectively, of Salmonella enterica serovar Senftenberg 775W and submitted to different ultrahigh pressure homogenization (UHPH) treatments at 150, 200, and 250 MPa. The inlet temperature of the samples was 6 degrees C. Counts of viable and injured Salmonella cells were obtained 2 h after the UHPH treatments and after 5, 10, 15, and 20 days of storage at 4 degrees C. The level of pressure applied influenced the lethality attained significantly (P < 0.05). In the samples with an initial load of approximately 7 log CFU/ml, the highest lethality value of 3.2 log CFU/ml was obtained at 250 MPa, and it is similar to those values reported in other surveys for thermal pasteurization with this same Salmonella strain. When the initial load was approximately 3 log CFU/ml, total inactivation was apparently obtained after the 250-MPa treatment (2.7 log CFU/ml). After 10 days of storage at 4 degrees C, Salmonella counts decreased in UHPH-treated samples, and colonies were not observed in tryptone soy agar and yeast extract medium. Nevertheless, presence of viable Salmonella cells was detected with the VIDAS Salmonella immunoassay method during the entire storage period. These results encourage further investigation of UHPH processing of liquid whole egg, assaying the possibility of using higher pressures and fluid inlet temperatures.

Outcomes of surgical treatment of diabetic foot osteomyelitis: a series of 185 patients with histopathological confirmation of bone involvement.

AIMS/HYPOTHESIS: We analysed the factors that determine the outcomes of surgical treatment of osteomyelitis of the foot in diabetic patients given early surgical treatment within 12 h of admission and treated with prioritisation of foot-sparing surgery and avoidance of amputation. METHODS: A consecutive series of 185 diabetic patients with foot osteomyelitis and histopathological confirmation of bone involvement were followed until healing, amputation or death. RESULTS: Probing to bone was positive in 175 cases (94.5%) and radiological signs of osteomyelitis were found in 157 cases (84.8%). Staphylococcus aureus was the organism isolated in the majority of cultures (51.3%), and in 35 cases (36.8%) it proved to be methicillin-resistant. The surgical treatment performed included 91 conservative surgical procedures, which were defined as those where no amputation of any part of the foot was undertaken (49.1%). A total of 94 patients received some degree of amputation, consisting of 79 foot-level (minor) amputations (42.4%) and 15 major amputations (8%). Five patients died during the perioperative period (2.7%). Histopathological analysis revealed 94 cases (50.8%) of acute osteomyelitis, 43 cases (23.2%) of chronic osteomyelitis, 45 cases (24.3%) of acute exacerbation of chronic osteomyelitis and three remaining cases (1.6%) designated as 'other'. The risks of failure in the case of conservative surgery were exposed bone, the presence of ischaemia and necrotising soft tissue infection. CONCLUSIONS/INTERPRETATION: Conservative surgery without local or high-level amputation is successful in almost half of the cases of diabetic foot osteomyelitis. Prospective trials should be undertaken to determine the relative roles of conservative surgery versus other approaches.

Hiperlucencia mediastínica radiológica en una crisis de asma / No disponible

El neumomediastino espontáneo o primario es una entidad poco frecuente, pero que en edad pediátrica se asocia con asiduidad a asma y sobre todo como complicación de una primera crisis asmática. La mayoría de veces se trata de un diagnóstico casual, aunque en caso de producir síntomas, éstos pueden ser muy variados, siendo los más frecuentes el dolor torácico y la disnea. El diagnóstico de sospecha se realiza en muchas ocasiones al palpar enfisema subcutáneo de localización cervical, aunque es la radiografía de tórax el diagnóstico de certeza. El curso es benigno y se resuelve con celeridad tras tratamiento sintomático

Primary or spontaneous pneumomediastinum is an uncommondisease. However, in the pediatric age, it is frequently associated with asthma and above all with complication of a first asthmatic episode. Usually it is found as a casual diagnosis, although when there are symptoms, these may be quite different, the most frequent being chest pain and dyspnea. The diagnosis suspicion is often made on palpating subcutaneous cervical emphysema, although the diagnostic certainty is obtained from the chest x-ray. It has a benign course and is resolved quickly after symptomatic treatment

Metahemoglobinemia en una lactante por consumo de puré vegetal / Infant methemoglobinemia after eating mixed vegetable puree

La cianosis es un signo clínico consistente en coloración azulada de piel y mucosas debida a un aumento de la hemoglobina reducida en los capilares, o menos frecuentemente, a la presencia de metahemoglobinemia (forma férrica de la hemoglobina) que puede ser ocasionada por contacto o ingesta de agentes oxidantes exógenos tóxicos como tintes de anilina, nitrobenceno, fármacos o compuestos nitrogenados de diferente procedencia, como son las verduras con alto contenido en nitratos (1 y 2). Presentamos el caso clínico de una lactante de 8 meses que fue traída a urgencias por presentar cianosis labial y de partes acras (manos y pies), sin otro tipo de sintomatología, tras la ingestión de un puré vegetal preparado y conservado a temperatura ambiente. La determinación de metahemoglobina fue del 22,8% mediante determinación por cooximetría. La evolución del lactante fue satisfactoria con tratamiento con oxígeno y observación durante 24 horas (AU)

Cyanosis is a clinical sign due to the presence of bluish coloration of skin and mucosae, caused by an increase in the reduced form of hemoglobin in the capillaries, or, less frequently, to the presence of methemoglobinemia (ferric form of hemoglobin). Its origin can be contact or ingestion of exogenous oxidating toxic agents like aniline dyes, nitrobencene, drugs or nitrogen compounds from different origin or vegetables with high nitrate content (1 and 2). We report a case of a 8-month-old female infant who was brought to the emergency room with no symptoms except cyanosis of the lips and acral areas (hands and feet) after the ingestion of a mixed vegetable puree, prepared and conserved at room temperature. Her methemoglobin level determined by cooximetry was 22.8%. She was treated during 24 hour with oxygen and observation and evolved satisfactorily (AU)

¿Nos puede matar el verano? / Can summer kill us?

El golpe de calor es una emergencia médica y causa tratable de fracaso multiorgánico (FMO). Aparece cuando el organismo pierde el control de la temperatura corporal por fracaso del sistema termorregulador, lo que causa un ascenso de la temperatura central hasta 41°C o más. La falta de regulación calórica produce una alteración de la función celular con daño en los tejidos muscular, cerebral, vascular, hepático y renal, y puede producir complicaciones graves como rabdomiolisis. Es fundamental el diagnóstico precoz ya que, si no se toman las medidas iniciales oportunas, la tasa de mortalidad es muy alta. Hay dos formas habituales de presentación, una relacionada con el ejercicio en ambientes calurosos y otra forma clásica, durante intensas olas de calor. El tratamiento se basa en el enfriamiento inmediato junto con el soporte de órganos y sistemas

Heat stroke is a medical emergency and treatable cause of multiorgan failure (MOF). It appears when the body losses control of body temperature due to thermoregulatory system failure, which causes an increase in central temperature to 41° or more. Lack of calorie regulation produces an alteration of cell function with injury to muscular, cerebral, vascular, hepatic and renal tissues, and can cause serious complications such as rhabdomyolysis. Early diagnosis is fundamental. If the adequate initial steps are not taken, mortality rate is very high. There are two common presentation forms, one related with exercise in hot environments, and another classical ones, during intense heat waves. Treatment is based on immediate cooling together with organ and system support

Response of two Salmonella enterica strains inoculated in model cheese treated with high hydrostatic pressure.

The aim of this work was to determine the response to high hydrostatic pressure and the ability for survival, recovery, and growth of 2 strains of Salmonella enterica (Salmonella enteritidis and Salmonella typhimurium) inoculated in a washed-curd model cheese produced with and without starter culture. Inoculated samples were treated at 300 and 400 MPa for 10 min at room temperature and analyzed after treatment and after 1, 7, and 15 d of storage at 12 degrees C to study the behavior of the Salmonella population. Cheese samples produced with starter culture and treated at 300 and 400 MPa showed maximum lethality; no significant differences in the baroresistant behavior of both strains were detected. Nevertheless, when starter culture was not present, the maximum lethality was only observed in cheese samples treated at 400 MPa, in the case of S. enteritidis. Ability to repair and grow was not observed in model cheese produced with starter culture and cell counts of treated samples decreased after 15 d of storage at 12 degrees C. In cheese produced without starter culture, Salmonella cells showed the ability to repair and grow during the storage period, reaching counts over 3 log(10) (cfu/mL) in both applied treatments and serotypes. These results suggest that high hydrostatic pressure treatments are effective to reduce Salmonella population in this type of cheese, but the presence of the starter culture affects the ability of this microorganism to repair and grow during the storage period.

Fate of Escherichia coli strains inoculated in model cheese elaborated with or without starter and treated by high hydrostatic pressure.

The aim of this research was to study high hydrostatic pressure inactivation of two strains of Escherichia coli (E. coli O59:H21 [CECT 405] and E. coli O157:H7 [CECT 5947]) inoculated in washed-curd model cheese elaborated with and without starter and the ability of these strains for survival, recovery, and growth. Samples were treated at 300, 400, and 500 MPa for 10 min at 20 degrees C and analyzed after the treatment and after 1, 7, and 15 days of storage at 8 degrees C to study the behavior of Escherichia populations. Cheeses elaborated with starter showed the maximum lethality at 400 and 500 MPa, and no significant differences in the baroresistant behavior of either strains were detected, except for E. coli O157:H7 at 400 MPa in cell counts obtained with thin agar layer method medium, where the decrease value was significantly lower. In cheese elaborated without starter, the highest decrease value was observed at 500 MPa, except for E. coli O59:H21 in cell counts obtained with selective culture medium, where the highest decrease value was also found at 400 MPa. The ability to repair and grow was not observed in model cheese elaborated with starter, as cell counts of treated samples decreased after 15 days of storage at 8 degrees C. By contrast, in cheese elaborated without starter, all pressurized samples showed the trend to repair and grow during the storage period in both strains. These results suggest that the presence of starter and low pH values are the main factors that control the ability of Escherichia strains inoculated in this type of cheese and treated by high hydrostatic pressure to recover and grow.