Genotypic and phenotypic characterization of lactic acid bacteria isolated from Italian ryegrass silage

Twenty-three lactic acid bacteria (LAB) isolated from three cultivars (Akiaoba, Nagahahikari and Tachiwase) of Italian ryegrass (Lolium multiflorum Lam.) silage were precisely characterized by a combination of phenotypic tests, genotypic 16S ribosomal DNA sequencing and rapid PCR-based analyses, focusing on their useful phenotypes for silage preparation as inoculants. We successfully identified both at the species and subspecies levels: phenotypically novel Lactococcus lactis subsp. lactis, Lactobacillus brevis, Lactobacillus coryniformis subsp. torquens, Lactobacillus curvatus, Lactobacillus plantarum subsp. plantarum, Lactobacillus sakei subsp. carnosus, Leuconostoc mesenteroides subsp. dextranicum and Pediococcus parvulus. This is the first report to elucidate the presence of Lactobacillus coryniformis ssp. torquens and Leuconostoc mesenteroides subsp. dextranicum in Italian ryegrass silages. Physiological and biochemical tests revealed that phenotypic characteristics are different among the different strains of the same species and subspecies, and that the isolates show unique and diverse phenotypes related to fermentation factors, such as available carbohydrates, optimal growth pH and temperature. These results suggest that, for various well-preserved silage preparations, the isolates may be useful in producing novel inoculants corresponding to their optimally climatic and ecological niches.     

Microflora in traditional starter cultures for fermented milk, hurunge, from Inner Mongolia, China

Microflora were investigated in traditional starter cultures for fermented milk, hurunge, which are used for fermented dairy products by nomadic families in the Inner Mongolia Autonomic Region, China. The acid‐forming bacteria and yeast counts ranged from 1.8 × 10⁵ to 5.3 × 10⁸ c.f.u./g and from 6.1 × 10⁵ to 3.2 × 10⁶ c.f.u./g, respectively. Sixty‐six strains of lactic acid bacteria and 30 strains of yeasts were isolated and identified from three hurunge samples collected from the nomadic families. Lactococcus raffinolactis was the most predominant lactococci isolated from these samples. The other lactococci were Lactococcus lactis ssp. lactis, Lactococcus lactis ssp. cremoris, and Leuconostoc mesenteroides ssp. cremoris. Two major lactobacilli strains, Lactobacillus plantarum and Lactobacillus casei, were identified. In addition, Lactobacillus kefiranofaciens, Lactobacillus acetotolerans, which grew in 11% acetic acid culture medium, and Lactobacillus homohiochii, which grew in the culture medium containing 16% ethanol, were also identified. The isolated yeast strains were identified as Candida kefyr, Saccharomyces cerevisiae, Kluyveromyces marxianus var. lactis, Candida krusei and Candida valida.     

The investigation of probiotic potential of lactic acid bacteria isolated from traditional Mongolian dairy products

The aims of this study were to investigate the diversity of lactic acid bacteria (LAB) isolated from traditional Mongolian dairy products, and to estimate the probiotic potential of the isolated strains. We collected 66 samples of the traditional Mongolian dairy products tarag (n = 45), airag (n = 7), aaruul (n = 8), byasulag (n = 1) and eezgii (n = 5), from which 543 LAB strains were isolated and identified based on 16S ribosomal DNA sequence. The predominant species of those products were Lactobacillus (L.) delbrueckii ssp. bulgaricus, L. helveticus, L. fermentum, L. delbrueckii ssp. lactis and Lactococcus lactis ssp. lactis. However, we could not detect any LAB strains from eezgii. All LAB isolates were screened for tolerance to low pH and to bile acid, gas production from glucose, and adherence to Caco‐2 cells. In vitro, we found 10 strains possess probiotic properties, and almost identified them as L. plantarum or L. paracasei subspecies, based on 16S ribosomal DNA and carbohydrate fermentation pattern. These strains were differentiated from each other individually by randomly amplified polymorphic DNA analysis. Additionally, it was notable that 6/10 strains were isolated from camel milk tarag from the Dornogovi province.     

Identification of lactic acid bacteria isolated from corn stovers

One hundred and twenty‐six strains were isolated from corn stover in Henan Province, China, of which 105 isolates were considered to be lactic acid bacteria (LAB) according to Gram‐positive, catalase‐negative and mainly metabolic lactic acid product. Analysis of the 16S ribosomal DNA sequence of 21 representative strains was used to confirm the presence of the predominant groups and to determine the phylogenetic affiliation of isolates. The sequences from the various LAB isolates showed high degrees of similarity to those of the GenBank type strains between 99.4% and 100%. The prevalent LAB, predominantly Lactobacillus (85.6%), consisted of L. plantarum (33.3%), L. pentosus (28.6%) and L. brevis (23.7%). Other LAB species as Leuconostoc lactis (4.8%), Weissella cibaria (4.8%) and Enterococcus mundtii (4.8%) also presented in corn stover. The present study is the first to fully document corn stover‐associated LAB involved in the silage fermentation. The identification results revealed LAB composition inhabiting corn stover and enabling the future design of appropriate inoculants aimed at improving the fermentation quality of silage.     

Isolating and evaluating lactic acid bacteria strains for effectiveness on silage quality at low temperatures on the Tibetan Plateau

Four lactic acid bacteria (LAB) strains isolated from straw silages on the Tibetan Plateau were characterized, and their effects on the fermentation quality of Italian ryegrass (Lolium multiflorum Lam.) at different temperatures (10°C, 15°C and 25°C) were studied. These LAB isolates were evaluated using the acids production ability test, morphological observation, Gram staining, physiological, biochemical and acid tolerance tests. All the isolates (M1, LM8, LO7 and LOG9) could grow at 5‐20°C, pH 3.5‐7.0 and NaCl (3.0%, 6.5%). Strains M1, LM8, LO7 and LOG9 were identified as Lactobacillus plantarum, L. coryniformis, Pediococcus pentosaceus and P. acidilactici, respectively, by sequencing 16S ribosomal DNA. The four isolates were added to Italian ryegrass for ensiling for 30 days at various temperatures. Compared with the corresponding control, inoculating with isolates M1, LM8 and LO7 could improve the silage quality of Italian ryegrass at low temperatures, indicated by significantly (P < 0.05) higher lactic acid (LA) contents and ratios of lactic acid/acetic acid (LA/AA), and significantly (P < 0.05) lower pH and ammonia nitrogen/total nitrogen (AN/TN). Compared with other isolates, LM8 performed better at 10°C and 15°C, indicated by the higher (P < 0.05) LA content and ratio of LA/AA, and the lower (P < 0.05) pH and AN/TN.     

Characterization and application of lactic acid bacteria for tropical silage preparation

Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid‐producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA‐DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small‐scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science     

Fermentation quality and aerobic stability of mulberry silage prepared with lactic acid bacteria and propionic acid

The objective of this study was to examine the fermentation quality and aerobic stability of mulberry (Morus alba L.) silage prepared with lactic acid bacteria (LAB) and propionic acid (PA). The selected LAB strains Lactobacillus (L.) plantarum LC365281 (L1) and L. brevis LC365282 (L2), and commercial inoculant strains L. plantarum Gaofuji (GF) and L. buchneri Fresh (FR), and PA were used as additives for silage preparation. Silage treatments were designed as control, L1, L2, GF, FR, PA, PA + L1, PA + L2, PA + GF, or PA + FR. After 30 days of ensiling, the fermentation quality of silages treated with PA + L1 was improved, with a lower (p < 0.05) pH and NH₃‐N content than those of other treatments. During the aerobic exposure, the PA + LAB‐treated silages displayed an aerobic stability with stable pH value and lactic acid content. The results confirm that L. plantarum L1 and PA were the best additive combination for ensiling mulberry.     

Fermentation quality and chemical composition of shrub silage treated with lactic acid bacteria inoculants and cellulase additives

Effects of lactic acid bacteria (LAB) inoculants and cellulase additives on fermentation quality and chemical compositions of shrub silages were studied by using a small‐scale fermentation system. Two LAB inoculants of Qingbao (Lactobacillus plantarum, Pediococcus acidilacticii, Lactobacillus casei and Clostridium phage) and Caihe (Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici) and a commercial cellulase made from Trichoderma reesei were used as additives for intermediate pea‐shrub, rush bushclover, arborescent ceratoides and shrubby silage preparation. The crude protein, neutral detergent fiber and water‐soluble carbohydrate contents of the four shrub materials were 10.1–14.2, 62.6–67.2 and 1.9–3.5% on a dry matter basis, respectively. All shrub silages had pH 3.40–4.43, ammonia‐N 0.1–0.2% g/kg and lactic acid 1.3–2.9% on a fresh matter basis. The silage quality of LAB‐inoculated silages did not have a greater effect than control silages, except shrubby silage preparation. Silages treated with the cellulase, the pH of rush bushclover and shrubby sweetvetch silage were significantly (P < 0.05) lower and the lactic acid content were significantly (P < 0.05) higher than the control silages. The results confirmed that shrub contained a relatively high content of crude protein; its silages can be preserved in good quality, and they are new potential resources for livestock feed.     

生物添加剂和含水率对紫花苜蓿和猫尾草青贮发酵品质的影响

试验旨在研究紫花苜蓿绿汁发酵液（AFGJ）、猫尾草绿汁发酵液（TFGJ）、乳酸菌制剂（LAB）、纤维素酶（CEL）及LAB＋CEL（MIX）对紫花苜蓿和猫尾草青贮发酵品质的影响。用紫花苜蓿和猫尾草各2种含水率（高、低）的原料制作了青贮。每个处理设2次重复，常温下贮存50d，开封后测定青贮的发酵品质。研究结果表明，添加剂对紫花苜蓿和猫尾草青贮的发酵品质有显著影响（P〈0．05），除AFGJ和TFGJ高含水率紫花苜蓿青贮外，各种添加剂均不同程度地改善了高、低含水率紫花苜蓿和猫尾草青贮的发酵品质；含水率对紫花苜蓿和猫尾草青贮的发酵品质有显著影响（P〈0．05），2种牧草均是低含水率优于高含水率青贮的发酵品质；添加剂和含水率对青贮的发酵品质有显著的交互作用（P〈0．05）。综合各项指标，在高、低含水率紫花苜蓿青贮中均以添加MIX效果最好；在高含水率猫尾草青贮中，添加TFGJ和AFGJ效果相近且优于其他添加剂；在低含水率猫尾草青贮中添加AFGJ、TFGJ、LAB和MIX效果相近，且优于添加CEL。结果表明，适当降低原料含水率并选择适宜的添加剂可调制优质青贮。     

Effects of inoculants and environmental temperature on fermentation quality and bacterial diversity of alfalfa silage

To investigate the effects of inoculants and environmental temperature on fermentation quality and bacterial diversity of alfalfa silage, first‐cut alfalfa was ensiled with or without two screened lactic acid bacteria (LAB) strains, Lactobacillus plantarum, LP, and Lactobacillus casei, LC. Each treatment was divided into three parts and stored at 20°C, 30°C, 40°C, respectively. After 60 days ensiling, fermentation characteristics were measured and bacterial diversity was investigated by 16S ribosomal RNA gene sequencing using Illumina MiSeq platform. LP and LC decreased pH, coliform bacteria counts and increased lactic acid content at 20°C, and the two strains decreased pH, ammonia‐N concentration, coliform bacteria counts at 30°C. When the environmental temperature was 40°C, silage treated with LC showed lower LAB and coliform bacteria counts and higher lactic acid content than the untreated and LP treated silages. Butyric acid mainly appeared in silages stored at 40°C. The relative abundance of Lactobacillus in alfalfa silages stored at 20°C and 30°C was highest and increased after LP and LC were added. Garciella was another dominant genus in silages stored at 40°C. In conclusion, LP and LC improved fermentation quality of alfalfa silage by increasing Lactobacillus proportions at 20°C and 30°C; ensiling alfalfa at 40°C was difficult because of Garciella.     

Effect of a potential probiotics Lactococcus garvieae B301 on the growth performance,immune parameters and caecum microflora of broiler chickens

In this study, a novel Lactococcus garvieae B301 was isolated from the intestinal tract of a healthy piglet. L. garvieae B301 was tolerant to acid pH, simulated gastric and small intestinal transit juices, indicating that it was capable of surviving in the gastrointestinal tract. L. garvieae B301 was safe and beneficial to broilers, as broiler chickens supplemented with L. garvieae B301 had lower diarrhoea incidence and mortality than the Control. Moreover, supplementation of broiler diets with L. garvieae B301 resulted in an increase in body weight and the number of caecum lactic acid bacteria and Bifidobacterium spp., and decrease in feed‐to‐gain ratio and the number of caecum coliforms. It also had a positive effect on the thymus index and bursa of Fabricius index and enhanced serum levels of immune globulins. All these results showed that L. garvieae B301 could enhance the growth performance of broiler chickens and improve their health. Thus, L. garvieae B301 could be a promising feed additive for broiler chickens.     

单一和复合乳酸菌添加剂对扁穗牛鞭草青贮品质的影响

利用实验室前期筛出的3株耐高温乳酸菌作为添加剂,探究单一和复合乳酸菌添加剂对高温高湿地区扁穗牛鞭草青贮品质的影响,进一步筛选出最佳的乳酸菌添加配比。试验设8个处理,分别为:1)戊糖片球菌(PP04);2)食窦魏斯氏菌(WC10);3)植物乳杆菌(LP694);4)M-1(PP04∶WC10=2∶1);5)M-2(PP04∶LP694=1∶2);6)M-3(WC10∶LP694=2∶1);7)M-4(PP04∶WC10∶LP694=2∶1∶1);8)CK(对照,无乳酸菌添加)。青贮60 d后,测定其发酵品质、营养成分及微生物组成,结果表明:1)处理组的NH3-N/TN显著低于对照组(P<0.05);2)处理组的乳酸、干物质含量和干物质回收率都显著高于对照组(P<0.05);3)处理组的pH值都小于4.2,其中,M-4处理组的pH值最低,为3.86;4)M-4处理组的微生物菌群组成和感官品质优于其他处理,乳酸含量最高,NH3-N/TN最低。因此,在扁穗牛鞭草青贮中添加单一或者复合乳酸菌均有利于调制出优质的扁穗牛鞭草青贮饲料,3种乳酸菌复合添加的M-4处理组改善青贮饲料的青贮品质的效果最佳,在生产实践中具有推广利用的潜质。     

Effect of microbial inoculants on the fermentation characteristics,nutritive value,and in vitro digestibility of various forages

This study was aimed to examine the characteristics of three isolated lactic acid bacteria (LAB) strains (HG24, M1, and TG1), and to evaluate their effects on fermentation characteristics, nutritive value, and in vitro digestibility in a variety of substrates. The experiments were conducted with Italian ryegrass (Lolium multiflorum Lam.), tall fescue (Festuca arundinacea Schred.), and oat (Avena sativa L.) in laboratory silo (1 L) for ensiling 30 days. All the treated silages had significantly (p < 0.05) higher in vitro dry matter digestibility (IVDMD), and lower undesirable microorganism populations compared with controls. In Italian ryegrass and oat silages, all the inoculants improved the fermentation quality, evidenced by distinctly (p < 0.05) higher LA contents and ratios of LA to acetic acid, and lower pH, butyric acid and ammonia nitrogen contents. Strains HG24 and commercial inoculant G (Lactobacillus plantarum MTD‐1) performed better than strains M1 and TG1, indicated by obviously (p < 0.05) higher LA contents, LAB numbers, and IVDMD. In tall fescue silage, all the isolated LAB strains (HG24, M1 and TG1) markedly (p < 0.05) increased IVDMD compared with strain G. Strain HG24 is recommended as a good LAB inoculant for ensiling in various forages.     

Silage fermentation and ruminal degradation of stylo prepared with lactic acid bacteria and cellulase

In order to improve the silage fermentation of stylo (Stylosanthes guianensis ) in tropical areas, stylo silages were prepared with commercial additives Lactobacillus plantarum Chikuso‐1 (CH 1), L. rhamnasus Snow Lact L (SN ), Acremonium cellulase (CE ) and their combination as SN +CE or CH 1 + CE , and the fermentation quality, chemical composition and ruminal degradation of these silages were studied. Stylo silages treated with lactic acid bacteria (LAB ) or cellulase, the pH value and NH ₃‐N ? total‐N were significantly (P  <  0.05) decreased while the ruminal degradability of dry matter (DM ), crude protein (CP ), neutral detergent fiber (aNDF om) and acid detergent fiber (ADF om) were significantly (P  <  0.05) increased compared to control. Compared to LAB or cellulase‐treated silages, the DM , CP contents and relative feed value (RFV ), and the ruminal degradability in LAB plus cellulase‐treated silages were significantly (P  <  0.05) higher, but the aNDF om content was significantly (P  <  0.05) lower. CH 1 + CE treatment was more effective in silage fermentation and ruminal degradation than SN +CE treatment. The results confirmed that LAB or LAB plus cellulase treatment could improve the fermentation quality, chemical composition and ruminal degradation of stylo silage. Moreover, the combined treatment with LAB and cellulase may have beneficial synergistic effects on ruminal degradation.     

Effects of ethanol,molasses and Lactobacillus plantarum on the fermentation quality,in vitro digestibility and aerobic stability of total mixed ration silages in the Tibetan plateau of China

In Tibet, it is common practice to make and relocate total mixed ration (TMR) silages before feeding due to the uneven distribution of forages temporally and spatially. This study was conducted to investigate the effects of Lactobacillus plantarum (L), molasses (M) or ethanol (E) on the fermentation quality and aerobic stability of local adaptive TMR silage. After 45 days of ensiling, pH and ammonia nitrogen in inoculated TMR silages were significantly lower than control. During the first 6 days of the aerobic exposure test, a small fluctuation in lactic acid concentration for all TMR silages was observed, and then silages with ethanol continued this trend, while lactic acid in silage without ethanol sharply decreased until the end of the aerobic exposure period. Meanwhile, pH gradually increased along the aerobic exposure; silages treated with ethanol showed lower pH after 9 days of aerobic exposure. The population of yeast gradually increased during 6 days of aerobic exposure, after that an accelerated rise was observed in TMR silages without ethanol. The combinational beneficial effect of L. plantarum and ethanol was found in combined addition of ethanol and Lactobacillus plantarum silages (EL), indicated by intermediate fermentation quality and higher aerobic stability.     

Effects of inoculants on the fermentation characteristics and in vitro digestibility of reed canary grass (Phalaris arundinacea L.) silage on the Qinghai‐Tibetan Plateau

To effectively use local available grass resources to cover the winter feed shortage on the Qinghai‐Tibetan Plateau, direct‐cut and wilted reed canary grass (RCG) silages were prepared by using a rolled‐bale system, and their ensiling characteristics and in vitro digestibility were studied. Silages were treated without (control) or with inoculants including LP (Lactobacillus plantarum), LPLB (L. plantarum, L. buchneri), and LPLBc (L. plantarum, L. buchneri, and cellulase), and were stored at ambient temperature (5.7–14.6°C) for 90 days. Compared with control, the inoculated silages increased (p < .05) lactic acid and acetic acid contents, and reduced (p < .05) final pH value and ammonia‐N ratio of total N. The highest WSC content (41.2 g/kg DM) occurred for LPLB‐inoculated silage, whereas LPLBc‐treated silage displayed the lowest contents of NDF (522.9 g/kg DM) and ADF (275.5 g/kg DM). In addition, LPLBc‐inoculated silage had the highest in vitro gas production (51.0 ml/g DM), in vitro DM digestibility (619.3 g/kg DM), and metabolic energy (9.6 kJ/kg DM). These results confirmed that treatments with inoculants at ensiling could improve silage fermentation and in vitro digestibility of RCG, and this could be a potential winter feed for animals on the Qinghai‐Tibetan Plateau.     

Characteristics of isolated lactic acid bacteria and their effectiveness to improve stylo (Stylosanthes guianensis Sw.) silage quality at various temperatures

Two lactic acid bacteria (LAB) strains, Pediococcus pentosaceus SC1 and Lactobacillus paraplantarum SC2 isolated from king grass silage, were characterized and their effectiveness to improve the silage fermentation quality of stylo (Stylosanthes guianensis Sw.) was studied. Strain SC1 was able to grow at a high temperature of 45°C, while SC2 did not. SC2 normally grew at a low pH of 4.0, while SC1 could not. These two strains and a commercial inoculant of LAB (L. plantarum, LP) were used as additives to stylo silage preparation at various temperatures (20°C, 30°C and 40°C). All LAB inoculants significantly (P < 0.05) reduced the pH value and ammonia-N content, and increased the ratio of lactic acid to acetic acid and quality score compared with the control. In addition, inoculating LAB strains markedly (P < 0.05) reduced butyric acid content at the temperatures of 30°C and 40°C. Compared to SC2 and LP strains, strain SC1 was the most effective for improving stylo silage quality at 20°C, indicated by the increase in lactic acid, ratio of lactic acid to acetic acid and quality score. At 30°C and 40°C, there were no significant differences among SC1, SC2 and LP treatments in pH values, contents of acetic acid, butyric acid and ammonia-N (P > 0.05).     

New lactic acid bacterial strains from traditional Mongolian fermented milk products have altered adhesion to porcine gastric mucin depending on the carbon source

Attachment of lactic acid bacteria to the mucosal surface of the gastrointestinal tract is a major property of probiotics. Here, we examined the ability of 21 lactic acid bacterial strains isolated from traditional fermented milk products in Mongolia to adhere to porcine gastric mucin in vitro. Higher attachment was observed with Lactobacillus delbrueckii subsp. bulgaricus strains 6‐8 and 8‐1 than with Lactobacillus rhamnosus GG (positive control). Lactococcus lactis subsp. cremoris strain 7‐1 adhered to mucin as effectively as did strain GG. Heat inactivation decreased the adhesive ability of strains 6‐8 and 8‐1 but did not affect strain 7‐1. The adhesion of strains 6‐8, 7‐1 and 8‐1 was significantly inhibited when the cells were pretreated with periodate and trypsin, indicating that proteinaceous and carbohydrate‐like cell surface compounds are involved in the adhesion of these strains. The adhesion of strain 7‐1 was affected by the type of carbohydrate present in the growth medium, being higher with fructose than with lactose, galactose or xylose as the carbon source. The sugar content of 7‐1 cells grown on various carbohydrates was negatively correlated with its adhesive ability. We provide new probiotic candidate strains and new information regarding carbohydrate preference that influences lactic acid bacterial adhesion to mucin.     

呼伦贝尔草原野生牧草青贮中优良乳酸菌的分离及鉴定

本研究对呼伦贝尔草原牧草青贮饲料中的乳酸菌进行了分离、鉴定,旨为筛选出优良乳酸菌菌株。以4个地区不同群落牧草的青贮饲料为试验材料,并采用传统微生物学鉴定方法及16S rRNA序列分析对分离得到的乳酸菌菌株进行了鉴定。菌株DME53为草乳杆菌,DMG108为干酪乳杆菌,DMC80为植物乳杆菌,DMG138为短乳杆菌,DMG139为戊糖片球菌;其中菌株DMG138为异型发酵乳酸菌,其余4株菌均为同型发酵乳酸菌。除DMG139在40和45 ℃下生长微弱外,其余菌株均在5~45 ℃不同温度条件下, 3.0%和6.5% NaCl培养液中良好生长。除菌株DME53在pH 3.0下不能生长外,其余菌株均在pH 3.0~8.0条件下良好生长或微弱生长。菌株DMC80具有产酸能力较强、发酵速率较快、耐酸、耐低温等特性,可作为制备适用于呼伦贝尔地区青贮饲料菌制剂的优良菌株。     

Comparative single‐dose pharmacokinetics of orally administered florfenicol in rainbow trout (Oncorhynchus mykiss,Walbaum, 1792) at health and experimental infection with Streptococcus iniae or Lactococcus garvieae

This study evaluates changes in the pharmacokinetic behavior of a single oral dose of florfenicol in rainbow trouts experimentally infected with Lactococcus garvieae or Streptococcus iniae. One hundred and fifty fish were randomly divided into three equal groups: 1—healthy fish, 2—fish inoculated with S. iniae (2.87 × 10⁷ CFU/ml, i.p.), and 3—fish inoculated with L. garvieae (6.8 × 10⁵ CFU/ml, i.p.). Florfenicol was administered to all groups at 15 mg/kg by oral gavage. Blood sampling was performed at 0, 2, 3, 6, 8, 12, 24, 48, 72, and 120 hr after drug administration to each group, and plasma concentration of florfenicol was assayed by HPLC method. The MICs of florfenicol were 1.2 μg/ml and 5 μg/ml against L. garviae and S. iniae, respectively. Healthy fish showed higher values for most of the PK/PD parameters as compared to fish infected with L. garvieae which was reversed in fish infected with S. iniae. Fish infected with L. garvieae showed decreased relative bioavailability accompanied by increased volume of distribution at steady‐state (Vd_ss) and total body clearance (Cl_B)_. Infection with S. iniae increased the peak concentration of drug after administration (Cmax) and decreased elimination half‐life (T_{1/2 β})_, central compartment volume (V_c), and Vd_ss. In conclusion, infection with these bacteria can affect the pharmacokinetic behavior of florfenicol in rainbow trouts as shown by decreased bioavailability and increased total body clearance and volume of distribution in L. garvieae infection and decreased volume of distribution accompanied by increased Cmax in S. iniae‐infected fish.