Apparent digestibility of protein, energy, and amino acids in some selected feed ingredients for pacu Piaractus mesopotamicus

The apparent digestibility coefficients (ADCs) of amino acids (AA), protein, and energy in six feed ingredients (Brazilian fish meal, soybean meal, corn gluten meal, alcoholic yeast, corn, and wheat bran) we evaluated for pacu juveniles. In general, all ingredients showed high digestibility values for all AA, and differences among ADCs of individual amino acid were detected ( P  < 0.01). Corn gluten, soybean, and fish meals had the highest ADCs of AA. The ADCs of protein in fish meal, yeast, and corn gluten meal should not be used as AA digestibility indicators, because those showed differences up to 6.7% between the ADCs of protein and AA. All ingredients had lower ADCs of energy than corn gluten meal ( P  < 0.01). Lysine was the first limiting amino acid in fish meal, corn gluten meal, wheat bran, and corn, and the second limiting amino acid in soybean meal, as methionine was the first limiting amino acid in soybean meal and yeast. However, the soybean meal was the best quality protein source, as it had the highest digestible essential amino acid index. This demonstrated that digestible amino acid values can be used to formulate practical diets for pacu, preventing potential deficiencies or excess that might cause environmental and economic losses.     

Dietary digestible lysine requirement and essential amino acid to lysine ratio for pacu Piaractus mesopotamicus

To determine the digestible lysine requirement for pacu juveniles, a dose–response feeding trial was carried out. The fish (8.66 ± 1.13 g) were fed six diets containing the digestible lysine levels: 6.8, 9.1, 11.4, 13.2, 16.1 and 19.6 g kg^?1 dry diet. The gradual increase of dietary digestible lysine levels from 6.8 to 13.2 g kg^?1 did not influence the average values of the parameters evaluated (P > 0.05). The increase of dietary digestible lysine level to 16.1 g kg^?1 significantly improved weight gain (WG), specific growth rate (SGR), protein productive value (PPV), protein efficiency rate (PER), and apparent feed conversion rate (FCR), but was not different from fish fed diets containing 19.6 g kg^?1 lysine. Fish fed diets containing 16.1 and 19.6 g kg^?1 digestible lysine showed lower body lipid contents than fish in the other treatments. The digestible lysine requirement as determined by the broken‐line model, based on average WG values, was 16.4 g kg^?1. The other essential amino acid requirements were estimated based on the ideal protein concept and the value determined for lysine.     

Co-Feeding of Pacu,Piaractus mesopotamicus Holmberg (1887), Larvae with Artemia Nauplii and a Microencapsulated Diet

Abstract

Aiming at a precocious substitution of live prey by artificial diet, a 20-day experiment with pacu, Piaractus mesopotamicuslarvae using co-feeding and abrupt weaning strategies was set up. At the end of the experiment, larvae fed Artemia showed the best results (P < 0.05) in weight, total length and biomass, compared with other treatments. Larvae fed exclusively a microencapsulated diet never ingested the diet. Diet ingestion in co-fed and abrupt-weaned larvae was low, but did increase during the experiment; however, Artemia influenced diet ingestion on co-fed larvae. Careful considerations should be given to diet processing and formulation to ensure survival and growth of larvae fed exclusively on prepared diets.     

Depletion study and estimation of withdrawal periods for florfenicol and florfenicol amine in pacu (Piaractus mesopotamicus)

The intensive production of farmed fish is at a global all‐time high, and the control of bacteria proliferation in fish farms requires the frequent use of antimicrobials. This practice raises important environmental concerns related to the emergence of antimicrobial‐resistant bacteria strains. Only a few antimicrobial drugs have been approved for use in aquaculture, one of which is florfenicol. This work studies the depletion and withdrawal period of florfenicol and its main metabolite, florfenicol amine, in pacu (Piaractus mesopotamicus), a neotropical characin widely farmed in the southern hemisphere. Juvenile pacu (average weight of 724 g) were stocked in a closed‐loop laboratory system with controlled water temperature (25.8°C), and were fed for 10 consecutive days with a diet containing an intended dose of 10 mg/florfenicol per kg bw. Muscle and skin tissues were collected at 1, 3, 6, 8, 10, 12 and 16 days post‐treatment, and florfenicol and florfenicol amine were quantified using a validated ultra‐high performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) method. The limits of quantitation for florfenicol and florfenicol amine were 10 ng/g in muscle and 50 ng/g in skin. Considering a maximum residue limit of 1000 ng/g for the sum of florfenicol and florfenicol amine in muscle with skin in natural proportions a withdrawal period of 5 days (water temperature 25.8°C) or 129 degree days was calculated on the basis of the upper limit of the one‐sided 95% confidence interval for the 99^th percentile derived from the residue depletion study.     

Growth and metabolism of pacu (Piaractus mesopotamicus Holmberg 1887) juveniles fed diets containing different protein, lipid and carbohydrate levels

To verify the potential of lipids and carbohydrates to spare dietary protein and to understand the intermediary metabolism of interaction of these nutrients in pacu juveniles, an experiment was carried out to evaluate pacu physiological and performance parameters. The experimental design was completely randomized with 12 treatments in a 2 × 2 × 3 factorial arrangement, consisting of diets containing two digestible protein levels (200 and 230 g kg⁻¹ PD), two lipid levels (40 and 80 g kg⁻¹) and three carbohydrate levels (410, 460 and 500 g kg⁻¹). Fish‐fed 230 g kg⁻¹ digestable protein (DP) showed increased glycaemia, decreased hepatic glycogen, as well as a smaller intake index and better feed conversion ratio. The higher dietary lipid level (80 g kg⁻¹) reduced protein intake and serum protein concentration, increased liver and body fat content, but did not affect growth. At a lipid level of 80 g kg⁻¹, the increase in dietary carbohydrate levels promoted greater weight gain (WG), crude protein intake (CPI) and better feed conversion ratio (FCR). For fish fed diets containing 40 g kg⁻¹ lipid, the best energy‐productive values (EPV) were obtained at 460 g kg⁻¹ carbohydrate. Increased levels of the main nutrients in the diets reduced the levels of serum triglycerides, while the increase in energy concentration increased the hepatosomatic (HSI) and glycaemia index values. Pacu used lipids as effectively as carbohydrates in the maximization of protein usage, as long as dietary protein was at a level of 230 g kg⁻¹ DP. The physiological parameters indicated that the best balance between the DP, dietary lipid and carbohydrate levels within the ranged this trial was obtained at 230, 40 and 460 g kg⁻¹, respectively, without lower growth.     

Effects of processing on physical characteristics of diets with distinct levels of carbohydrates and lipids: the outcomes on the growth of pacu (Piaractus mesopotamicus)

The use efficiency and feed conversion of extruded and pelletized diets were compared. Eight isoproteic diets (220 g kg⁻¹ digestible protein) were assayed for 90 days in a 2 × 2 × 2 multifactorial design with two carbohydrate levels (400 and 500 g kg⁻¹), two lipids levels (40 and 80 g kg⁻¹) and two diet processing (pelletization and extrusion) with three repetitions. The growth of Piaractus mesopotamicus fed with these diets and the quality control indices of diets were gauged. The density of extruded diets was lower as carbohydrate level was 400 g kg⁻¹ and lipid 40 g kg⁻¹. The interaction carbohydrate and diet processing presented higher leaching value for low carbohydrate level in extruded diet. Fish fed with extruded diets presented the best feed conversion and protein efficiency ratio. When high levels of carbohydrate and lipid are combined, the weight gain is impaired. The interaction between diet processing diet and lipid levels resulted in the best fish performance when pelletized diets with 40 g kg⁻¹ lipid or extruded diets with 80 g kg⁻¹ lipid were considered. The protein efficiency ratio increased with the increment of carbohydrates in the pelletized diets. The fish show low tolerance to lipids and a preference for carbohydrate when the lipid productive values are taken into account.     

Digestibility and performance of pacu (Piaractus mesopotamicus) juveniles — fed diets containing different protein, lipid and carbohydrate levels

Due to lack of information on the use of non‐protein energy sources in diets for pacu (Piaractus mesopotamicus), a 2 × 2 × 3 factorial experiment was conducted to evaluate the performance and digestibility of 12 diets containing approximately two crude protein (CP; 220 and 250 g kg⁻¹), two lipid (40 and 80 g kg⁻¹) and three carbohydrate levels (410, 460 and 500 g kg⁻¹). The pacu juveniles‐fed diets containing 220 g kg⁻¹ CP did not respond (P > 0.05) to increased dietary lipid and carbohydrate levels, but the fish‐fed diets containing 250 g kg⁻¹ CP showed a better feed conversion ratio. There were interactions in weight gain (WG), specific growth rate (SGR), crude protein intake (CPI) and feed conversion rate (FCR) dependent on dietary carbohydrate and lipid levels, showing positive effects of increasing carbohydrate levels only for fish‐fed diets containing 80 g kg⁻¹ lipid level. However, when the diets contained 40 g kg⁻¹ lipid, the best energy productive value (EPV) results were obtained at 460 g kg⁻¹ carbohydrate. A higher usage of lipids (80 g kg⁻¹) reduced CPI and was detrimental to protein [apparent digestibility coefficient (ADC)_CP] and energy (ADC_GE), but did not affect growth. The ADC_GE improved proportionally as dietary carbohydrate levels increased (P < 0.05), increasing the concentration of digestible energy. In addition, the WG, CPI, ADC_GE results showed best use of the energy from carbohydrates when dietary protein level was 250 g kg⁻¹ CP. The utilization of 250 g kg⁻¹ CP in feeds for juvenile pacu for optimal growth is suggested. Therefore, the optimum dietary lipid and carbohydrate levels depend on their combinations. It can be stated that pacu uses carbohydrates as effectively as lipids in the maximization of protein usage, as long as it is not lower than 250 g kg⁻¹ CP or approximately 230 g kg⁻¹ digestible protein.     

Growth and haematology of pacu, Piaractus mesopotamicus, fed diets with varying protein to energy ratio

Haematopoiesis and blood cells' functions can be influenced by dietary concentration of nutrients. This paper studied the effects of dietary protein:energy ratio on the growth and haematology of pacu, Piaractus mesopotamicus . Fingerling pacu (15.5±0.4 g) were fed twice a day for 10 weeks until apparent saciety with diets containing 220, 260, 300, 340 or 380 g kg⁻¹ crude protein (CP) and 10.88, 11.72, 12.55, 13.39, 14.22 MJ kg⁻¹ digestible energy (DE) in a totally randomized experimental design, 5 × 5 factorial scheme ( n =3). Weight gain and specific growth rate were affected ( P <0.05) by protein level only. Protein efficiency ratio decreased ( P <0.05) with increasing dietary protein at all levels of dietary energy. Daily feed intake decreased ( P <0.05) with increasing dietary energy. Mean corpuscular haemoglobin concentration was affected ( P <0.05) by DE and interaction between dietary CP and DE. Total plasma protein increased ( P <0.05) with dietary protein and energy levels. Plasma glucose decreased ( P <0.05) with increasing dietary protein. The CP requirement and optimum protein:energy ratio for weight gain of pacu fingerlings, determined using broken-line model, were 271 g kg⁻¹ and 22.18 g CP MJ⁻¹ DE respectively. All dietary CP and DE levels studied did not pose damages to fish health.     

Growth,nitrogen gain and indispensable amino acid retention of pacu (Piaractus mesopotamicus,Holmberg 1887) fed different brewers yeast (Saccharomyces cerevisiae) levels

A feeding‐and‐digestibility trials were carried out to evaluate the efficacy of replacing fishmeal with brewers yeast Saccharomyces cerevisiae in diets of pacu, Piaractus mesopotamicus, juveniles. The feeding trial was conducted during 54 days with 450 fish (26.6 ± 1.7 g) testing six isonitrogenous (270 g kg^?1 crude protein) and isoenergetic (19 MJ kg^?1 crude energy) diets, with increasing yeast level to replace 0 (control), 30, 35, 50, 70 or 100% of dietary fishmeal. Growth performance and feed utilization increased with increasing dietary yeast level until 50% fishmeal replacement. Protein retention efficiency was higher in fish fed 35 and 50%. Protein digestibility and the fillet hue (the red/green chromaticity) were not significantly different among all treatments. Nitrogen gains were significantly improved in fish fed 35% replacement diet compared to fish fed the control diet. The retentions of indispensable amino acids tended to increase with increasing dietary yeast levels, with maximum retention at 35–50%. On the basis of our results, replacing 50% fishmeal by yeast in pacu diets successfully improved feed efficiency and growth performance, and reduced nitrogen losses, thereby reducing the nitrogen outputs from fish farms.     

Growth performance and body composition of pacu Piaractus mesopotamicus (Holmberg 1887) in response to dietary protein and energy levels

Improper dietary protein and energy levels and their ratio will lead to increased fish production cost. This work evaluated effects of dietary protein : energy ratio on growth and body composition of pacu, Piaractus mesopotamicus. Fingerling pacu (15.5 ± 0.4 g) were fed twice a day for 10 weeks until apparent satiation with diets containing 220, 260, 300, 340 or 380 g kg^?1 crude protein (CP) and 10.9, 11.7, 12.6, 13.4 or 14.2 MJ kg^?1 digestible energy (DE) in a totally randomized experimental design, 5 × 5 factorial scheme (n = 3). Weight gain, specific growth rate increased and feed conversion ratio (FCR) decreased significantly (P < 0.05) when CP increased from 220 to 271, 268 and 281 g kg^?1 respectively. Pacu was able to adjust feed consumption in a wide range of dietary DE concentration. Fish fed 260 CP diets showed best (P < 0.05) protein efficiency ratio and FCR with 11.7–12.6 MJ kg^?1; but for the 380 CP‐diets group, significant differences were observed only at 14.2 MJ kg^?1 dietary energy level, suggesting that pacu favours protein as energy source. DE was the chief influence on whole body chemical composition. Minimum dietary protein requirement of pacu is 270 g kg^?1, with an optimum CP : DE of 22.2 g MJ^?1.     

Response of juvenile pacu (Piaractus mesopotamicus Holmberg, 1887) to balanced digestible protein

This 45‐day work aimed to determine the response of juvenile pacu (Piaractus mesopotamicus) to balanced digestible protein (BDP) and to use these responses to determine whether the optimum economic levels of BDP would differ depending on the form in which the fish is sold. Six isoenergetic diets containing 163, 201, 238, 272, 315 and 348 g/kg BDP (dry‐matter basis) were prepared through the serial dilutions of a high‐protein diet with the low‐protein diet. Fish (initial average body weight, 10.82 ± 0.14 g) were fed with respective experimental diets three times a day until apparent satiation. The optimum biological level of BDP was calculated as 326 g/kg (dry‐matter basis) by the quadratic regression model for maximum body weight gain of juvenile pacu. To maximize economic returns (US$/kg) for different end products, the optimum economic levels of BDP were calculated as 311, 317 and 319 g/kg (dry‐matter basis) by an economic model for whole‐body, eviscerated and sliced juvenile pacu respectively. This finding revealed that obtaining maximum biological performance of fish in the case of high feed costs or low prices of the end product in consumer market would substantially reduce the economic returns.     

The effect of dietary protein level on pond water quality and feed utilization efficiency of pacúPiaractus mesopotamicus (Holmberg, 1887)

The effect of supplemental feeds with different levels of crude protein on pond water quality and food utilization efficiency by pacú (Piaractus mesopotamicus) was evaluated in a semi‐intensive culture system. Fish were stocked at a density of one individual per m², raised for 299 days, and fed isocaloric diets containing soybean meal, blood and bone meal, and yellow maize. Dietary treatments consisted of diets containing 25%, 35%, and 45% crude protein. Ponds receiving the two highest protein levels showed significantly higher values of alkalinity, conductivity and nitrites. High dietary protein diets resulted in no significant improvements in final weight (336.4±77.2, 308.2±92.6 and 368.4±82.2 g, mean±standard deviation, for 25%, 35% and 45% protein levels respectively). Moreover, fish carcass composition was not significantly different among treatments. Instead, significant augmentations in feed conversion ratio (1.36, 1.54 and 1.73 respectively) and reductions in protein retention (35.7%, 33.5% and 29.0% respectively) occurred as protein levels increased. Results indicate that at the lowest protein level, pacú growth rates and carcass composition were similar to the other experimental treatments, with a significant improvement in pond water quality and feed utilization efficiency.     

Carbohydrate tolerance in the fruit‐eating fish Piaractus mesopotamicus (Holmberg, 1887)

Some fish species have a limited ability to metabolize dietary carbohydrates. An important tool for understanding carbohydrate metabolism is the application of the glucose tolerance test, which can be performed orally or intraperitoneally. To evaluate carbohydrate tolerance in the fruit‐eating fish pacu, two experiments were performed, one with oral administration by gavage of three carbohydrate types (glucose, fructose and starch, 2.0 g/kg body weight (BW)) and the other with intraperitoneal injection (IP) of glucose (500 mg/kg BW). Oral glucose resulted in an increase in plasma glucose 2 hr later with the peak at 4 hr (8.30 mmol/L), and return to baseline between 6 and 12 hr; starch administration promoted a peak after 4 hr (7.70 mmol/L), returning to the baseline at 6 hr. The administration of fructose promoted a moderate peak after 2 hr (5.71 mmol/L), and return to baseline for the time points that followed. Elevated serum cholesterol levels were observed 2 and 24 hr after administration of glucose and starch. Hepatic glycogen levels increased within 24 hr, regardless of the type of carbohydrate administered. IP glucose load resulted in a peak of plasma glucose 3 hr post injection (6.91 mmol/L), returning to baseline 6 hr later. There was a reduction in the concentration of triglycerides at 24 hr. The results demonstrate that pacu metabolize both oral (glucose or starch) and intraperitoneal (glucose) carbohydrate loads after 6 hr, suggesting good ability to deal with dietary carbohydrates.     

Whole-body amino acid pattern of juvenile,preadult, and adult pacu,Piaractus mesopotamicus,with an estimation of its dietary essential amino acid requirements

In the present study, juvenile (live body weight, 54.3 ± 8.2 g), preadult (live body weight, 822.5 ± 33.9 g), and adult (live body weight, 1,562.8 ± 41.8 g) pacu, Piaractus mesopotamicus, were used to estimate their dietary essential amino acid (EAA) requirements using the whole-body amino acid (AA) pattern. The results showed that whole-body moisture, crude protein, total lipid, and ash contents expressed on a wet weight basis (%) were significantly different among the studied growth phases. No significant differences were observed in the dietary EAA requirements estimated for the three growth phases of pacu. These dietary EAA requirements were found to be different than those previously estimated for the same fish through its muscle AA pattern. Based on whole-body EAA to total EAA ratios {A/E ratios; [(each EAA/total EAA) × 1,000]}, EAA requirements were estimated to be histidine (0.42%), arginine (1.36%), threonine (0.82%), valine (0.90%), methionine (0.45%), isoleucine (0.83%), leucine (1.29%), phenylalanine (0.74%), lysine (1.64%), and tryptophan (0.14%) for pacu. These estimated requirements may serve as a reference line in the formulation of practical and experimental diets until dose–response-based optimum EAA requirements are available for pacu.     

Essential oils to control ichthyophthiriasis in pacu,Piaractus mesopotamicus (Holmberg): special emphasis on treatment with Melaleuca alternifolia

In vitro effect of the Melaleuca alternifolia, Lavandula angustifolia and Mentha piperita essential oils (EOs) against Ichthyophthirius multifiliis and in vivo effect of M. alternifolia for treating ichthyophthiriasis in one of the most important South American fish, Piaractus mesopotamicus (Holmberg), were evaluated. The in vitro test consisted of three EOs, each at concentrations of 57 μL L^?1, 114 μL L ^?1, 227 μL L^?1 and 455 μL L ^?1, which were assessed once an hour for 4 h in microtitre plates (96 wells). The in vitro results demonstrated that all tested EOs showed a cytotoxic effect against I. multifiliis compared to control groups (P < 0.05). The in vivo treatment for white spot disease was performed in a bath for 2 h day^?1 for 5 days using the M. alternifolia EO (50 μL L ^?1). In this study, 53.33% of the fish severely infected by I. multifiliis survived after the treatment with M. alternifolia (50 μL L ^?1) and the parasitological analysis has shown an efficacy of nearly 100% in the skin and gills, while all the fish in the control group died. Furthermore, the potential positive effect of M. alternifolia EO against two emergent opportunistic bacteria in South America Edwardsiella tarda and Citrobacter freundii was discussed.     

Acute toxicity of ammonia in Pacu fish (Piaractus mesopotamicus,Holmberg, 1887) at different temperatures levels

Piaractus mesopotamicus juveniles (total length 12 ± 0.5 mm) were exposed to different concentrations of ammonia‐N (un‐ionized plus ionized ammonia as nitrogen), using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7. The 24, 48, 72, 96 h LC₅₀ values of ammonia‐N in P. mesopotamicus juveniles were 5.32, 4.19, 3.79 and 2.85 mg L^?1 at 15°C; 4.81, 3.97, 3.25 and 2.50 mg L^?1 at 20°C; and 4.16, 3.79, 2.58 and 1.97 mg L^?1 at 25°C respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃‐N (un‐ionized ammonia as nitrogen) were 0.018, 0.014, 0.013, 0.009 mg L^?1 at 15°C temperature; 0.023, 0.019, 0.016 and 0.012 mg L^?1 at 20°C; 0.029, 0.026, 0.018 and 0.014 mg L^?1 at 25°C. The temperature increase from 15 to 25°C caused an increase of ammonia‐N susceptibility by 21.80%, 9.55%, 31.92% and 30.87%, after 24, 48, 72 and 96 h exposure respectively. Furthermore, we found that exposure of fish to ammonia‐N caused an elevation in total haemoglobin and blood glucose with an increase of 2 mg L^?1 concentration. Ammonia levels tolerated, especially in different temperatures levels, have important implications for the management of aquaculture.     

Stable carbon (δ13C) and nitrogen (δ15N) isotopes as natural indicators of live and dry food in Piaractus mesopotamicus (Holmberg, 1887) larval tissue

This study proposed the use of the stable isotope technique to track the type of food utilized by pacu Piaractus mesopotamicus larvae during their development, and to identify the moment when the larvae start using nutrients from the dry diet by retaining its carbon and nitrogen atoms in their body tissues. Five‐day‐old pacu larvae at the onset of exogenous feeding were fed Artemia nauplii or formulated diet exclusively; nauplii+formulated diet during the entire period; or were weaned from nauplii to a dry diet after 3, 6 or 12 days after the first feeding. δ¹³C and δ¹⁵N values for Artemia nauplii were ?15.1‰ and 4.7‰, respectively, and ?25.0‰ and 7.4‰ for the dry diet. The initial isotopic composition of the larval tissue was ?20.2‰ and 9.5‰ for δ¹³C and δ¹⁵N respectively. Later, at the end of a 42‐day feeding period, larvae fed Artemia nauplii alone reached values of ?12.7‰ and 7.0‰ for δ¹³C and δ¹⁵N respectively. Larvae that received the formulated diet alone showed values of ?22.7‰ for δ¹³C and 9.6‰ for δ¹⁵N. The stable isotope technique was precise, and the time at which the larvae utilized Artemia nauplii, and later dry diet as a food source could be clearly defined.     

Fasting and refeeding lead to more efficient growth in lean pacu (Piaractus mesopotamicus)

We evaluated whether body fat content affects the energetic metabolism and growth in pacu submitted to daily feeding, fasting and refeeding. For 15 days, fish were fed different diets to obtain lean and fat conditions, and then subjected, for 20 days to: (1) continuously feeding (control), or (2) fasting for 15 days and refeeding for 5 days. Blood (glucose, triglycerides, cholesterol, non‐esterified fatty acids and total protein) and tissue (liver lipid and glycogen, muscle lipid and mesenteric fat) metabolic indicators, and growth performance parameters (weight gain, specific growth rate, daily feed intake and feed conversion ratio) were measured. Fasting led both lean and fat pacu to make notable use of their energy reserves, through glycogenolysis and lipolysis, reflected in reduced blood glucose and triglycerides, liver glycogen and muscle lipid levels. Lipolysis was confirmed by the high levels of non‐esterified fatty acids, especially in fat pacu. Refeeding led to higher plasma glucose and liver lipid in lean fish. Muscle fat increased in fat fish but was not restored in lean fish, while mesenteric fat index (MFI) remained the same in fat fish and increased in lean fish. Although refeeding occurred only for 5 days, lean fish grew more and were more efficient at utilizing food (higher weight gain and better feed conversion ratio). In conclusion, our results suggest that fat pacu have higher glycogenic and lipogenic abilities, and the higher deposition of lipids in fish does not mean higher availability of energy for growth when compensatory growth is stimulated by refeeding after fasting.